Novel Nucleoside Derivatives

ABSTRACT

Compounds of Formulae I-XVI, stereoisomers, and pharmaceutically acceptable salts or prodrugs thereof, their preparation, and their uses for the treatment of viral diseases including hepatitis C viral infection, cancer, diabetes, and other diseases are described: formula (I).

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention is directed towards novel 2′,3′-cycliccarbonate-containing nucleosides and their derivatives, including5′-monophosphate derivatives, their preparation and their uses. Morespecifically, the novel compounds are useful in the treatment of viralinfections, including hepatitis C viral infections, as well as cancerand other diseases and disorders for which treatment with nucleosidederivatives is useful or efficacious.

2. Background Art

The following description of the background of the invention is providedto aid in understanding the invention, but is not admitted to be, or todescribe, prior art to the invention. All publications are incorporatedby reference in their entirety.

Hepatitis C is a viral disease that causes inflammation of the liverthat may lead to cirrhosis, primary liver cancer and other long-termcomplications. Nucleosides are a well-recognized class of compoundsshown to be effective against a variety of viral infections, includingthose caused by hepatitis B virus (HBV), hepatitis C virus (HCV), humanimmunodeficiency virus (HV), and herpes virus.

Nucleosides are generally effective as antiviral agents followingconversion of the nucleoside to the corresponding nucleoside5′-triphosphate (NTP). Conversion occurs inside cells through the actionof various intracellular kinases. The first step, i.e., conversion ofthe nucleoside to the 5′-monophosphate (NMP), is generally the slow stepand involves a nucleoside kinase, which is encoded by either the virusor host. Conversion of the NMP to the NTP is generally catalyzed by hostnucleotide kinases. The NTP interferes with viral replication throughinhibition of viral polymerases and/or via incorporation into a growingstrand of DNA or RNA followed by chain termination.

Use of nucleosides to treat viral liver infections is often complicatedby one of two problems. In some cases, the desired nucleoside is a goodkinase substrate and accordingly produces NTP in the liver as well asother cells and tissues throughout the body. Since NTP production isoften associated with toxicity, efficacy can be limited by extrahepatictoxicities. In other cases, the desired nucleoside is a poor kinasesubstrate so is not efficiently converted into the NMP and ultimatelyinto the NTP.

For instance, U.S. Pat. No. 6,312,662 discloses the use of certainphosphate prodrugs for the liver-specific delivery of various drugsincluding nucleosides for the treatment of patients with liver diseasessuch as hepatitis C, hepatitis B and hepatocellular carcinoma.

SUMMARY OF THE INVENTION

The present invention is directed towards novel nucleoside derivatives,their preparation and their uses for the treatment of diseases anddisorders responsive to a pharmaceutical composition comprising anucleoside as an active pharmaceutical ingredient, including, e.g.,viral infections and cancer.

In some aspects, the invention concerns 2′,3′-cyclic carbonatenucleoside and nucleotide compounds and their derivatives and prodrugsthereof. The invention further relates to the treatment of diseases ordisorders using the disclosed 2′,3′-cyclic carbonate nucleoside ornucleotide compounds, derivatives, or prodrugs thereof.

Thus, in some aspects, the present invention relates to a compound ofFormula I, or an isomer, solvate, hydrate, prodrug or pharmaceuticallyacceptable salt thereof:

wherein:

X′, Y, R¹⁹, R¹⁸, R¹⁷, R¹⁶, R¹⁵, B, Z′, and Z″ are as defined below.

The present invention is also directed to a compound of Formula II:

or an isomer, solvate, hydrate, prodrug, or pharmaceutically acceptablesalt thereof, wherein X′, Y, R¹⁹, R¹⁸, R¹⁷, R¹⁶, R¹⁵, B, V, Z, W, and W′are as defined below.

The present invention is further directed to a pharmaceuticalcomposition comprising a compound of the present invention and apharmaceutically acceptable excipient or carrier.

The present invention is further directed to a method of treating adisease or disorder responsive to treatment with a pharmaceuticalcomposition comprising a nucleoside derivative as an activepharmaceutical ingredient.

The present invention is further directed to a method of treating aviral infection in a patient in need thereof, the method comprisingadministering to the patient a therapeutically effective amount of acompound of the present invention.

The present invention is further directed to a method of treating an HCVor HBV viral infection in a patient in need thereof, the methodcomprising administering to the patient a therapeutically effectiveamount of a compound of the present invention.

The present invention is also directed to a method of inhibiting viralreplication in a patient in need thereof, the method comprisingadministering to the patient a therapeutically effective amount of acompound of the present invention.

The present invention is also directed a method of treating cancer in apatient in need thereof, the method comprising administering to thepatient a therapeutically effective amount of a compound of the presentinvention.

The present invention is also directed a method of treating a plateletdisorder or diabetes in a patient in need thereof, the method comprisingadministering to the patient a therapeutically effective amount of acompound of the present invention, wherein said compound is a P2receptor antagonist.

The present invention is also directed a method of treating diabetes orcardiovascular disease in a patient in need thereof, the methodcomprising administering to the patient a therapeutically effectiveamount of a compound of the present invention, wherein said compoundbinds an adenosine receptor.

The present invention is also directed a method of treating inflammationor a CNS disorder in a patient in need thereof, the method comprisingadministering to the patient a therapeutically effective amount of acompound of the present invention, wherein said compound acts as anadenosine analogue.

DEFINITIONS

In accordance with the present invention and as used herein, thefollowing terms are defined with the following meanings, unlessexplicitly stated otherwise.

The term “alkyl” refers to saturated aliphatic groups includingstraight-chain, branched chain and cyclic groups, up to and including 12carbon atoms, or, more preferably, up to and including 10 carbon atoms,or up to and including 6 carbon atoms. Suitable alkyl groups includemethyl, ethyl, n-propyl, isopropyl, and cyclopropyl. The alkyl may beoptionally substituted with 1-3 substituents.

The term “aryl” refers to aromatic groups which have 5-14 ring atoms,and at least one ring having a conjugated pi electron system andincludes carbocyclic aryl, heterocyclic aryl and biaryl groups, all ofwhich may be optionally substituted. The aryl group may be optionallysubstituted with 1-6 substituents.

Carbocyclic aryl groups are groups which have 6-14 ring atoms whereinthe ring atoms on the aromatic ring are carbon atoms. Carbocyclic arylgroups include monocyclic carbocyclic aryl groups and polycyclic orfused compounds such as optionally substituted naphthyl groups.

Heterocyclic aryl or heteroaryl groups are groups which have 5-14 ringatoms wherein 1 to 4 heteroatoms are ring atoms in the aromatic ring andthe remainder of the ring atoms being carbon atoms. Suitable heteroatomsinclude oxygen, sulfur, and nitrogen. Suitable heteroaryl groups includefuranyl, thienyl, pyridyl, pyrrolyl, N-lower alkyl pyrrolyl,pyridyl-N-oxide, pyrimidyl, pyrazinyl, imidazolyl, indolyl and the like,all optionally substituted.

The term “monocyclic aryl” refers to aromatic groups which have 5-6 ringatoms and includes carbocyclic aryl and heterocyclic aryl. Suitable arylgroups include phenyl, furanyl, pyridyl, and thienyl. Aryl groups may besubstituted.

The term “monocyclic heteroaryl” refers to aromatic groups which have5-6 ring atoms wherein 1 to 4 heteroatoms are ring atoms in the aromaticring and the remainder of the ring atoms being carbon atoms. Suitableheteroatoms include oxygen, sulfur, and nitrogen.

The term “biaryl” represents aryl groups which have 5-14 atomscontaining more than one aromatic ring including both fused ring systemsand aryl groups substituted with other aryl groups. Such groups may beoptionally substituted. Suitable biaryl groups include naphthyl andbiphenyl.

The term “optionally substituted” or “substituted” includes groupssubstituted by one to four substituents, independently selected fromlower alkyl, lower aryl, lower aralkyl, lower cyclic alkyl, lowerheterocycloalkyl, hydroxy, lower alkoxy, lower aryloxy, perhaloalkoxy,aralkoxy, lower heteroaryl, lower heteroaryloxy, lower heteroarylalkyl,lower heteroaralkoxy, azido, amino, halogen, lower alkylthio, oxo, loweracyl, lower acylalkyl, lower carboxy esters, sulfonyl, sulfonylamido,carboxyl, -carboxamido, nitro, lower acyloxy, lower aminoalkyl, loweralkylamino, lower alkylaminoaryl, lower alkylaryl, loweralkylaminoalkyl, lower alkoxyaryl, lower arylamino, lower aralkylamino,lower alkylsulfonyl, lower -carboxamidoalkylaryl, lower-carboxamidoaryl,lower hydroxyalkyl, lower haloalkyl, lower alkylaminoalkylcarboxy-,lower aminocarboxamidoalkyl-, cyano, lower alkoxyalkyl, lowerperhaloalkyl, and lower arylalkyloxyalkyl. “Substituted aryl” and“substituted heteroaryl” refers to aryl and heteroaryl groupssubstituted with 1-6 of the substituents listed above. Preferredsubstituents are those selected from the group consisting of loweralkyl, lower alkoxy, lower perhaloalkyl, halogen, hydroxy, cyano, andamino.

The term “-aralkyl” refers to an alkylene group substituted with an arylgroup. Suitable aralkyl groups include benzyl, picolyl, and the like,and may be optionally substituted. The aryl portion may have 5-14 ringatoms and the allyl portion may have up to and including 10 carbonatoms. “Heteroarylalkyl” refers to an alkylene group substituted with aheteroaryl group.

The term “alkylaryl-” refers to an aryl group substituted with an alkylgroup. “Lower alkylaryl-” refers to such groups where alkyl is loweralkyl. The aryl portion may have 5-14 ring atoms and the alkyl portionmay have up to and including 10 carbon atoms. The term “lower” referredto herein in connection with organic radicals or compounds respectivelydefines such as with up to and including 10, in one aspect up to andincluding 6, and in another aspect one to four carbon atoms. Such groupsmay be straight chain, branched, or cyclic.

The term “cyclic alkyl” or “cycloalkyl” refers to alkyl groups that arecyclic of 3 to 10 carbon atoms, and in one aspect are 3 to 6 carbonatoms. Suitable cyclic groups include norbornyl and cyclopropyl. Suchgroups may be substituted.

The term “heterocyclic,” “heterocyclic alkyl” or “heterocycloalkyl”refer to cyclic groups of 3 to 10 atoms, and in one aspect are 3 to 6atoms, containing at least one heteroatom, in a further aspect are 1 to3 heteroatoms. Suitable heteroatoms include oxygen, sulfur, andnitrogen. Heterocyclic groups may be attached through a nitrogen orthrough a carbon atom in the ring. The heterocyclic alkyl groups includeunsaturated cyclic, fused cyclic and spirocyclic groups. Suitableheterocyclic groups include pyrrolidinyl, morpholino, morpholinoethyl,and pyridyl.

The terms “arylamino” (a), and “aralkylamino” (b), respectively, referto the group —NRR′ wherein respectively, (a) R is aryl and R′ ishydrogen, alkyl, aralkyl, heterocycloalkyl, or aryl, and (b) R isaralkyl and R′ is hydrogen, aralkyl, aryl, alkyl or heterocycloalkyl.

The term “acyl” refers to —C(O)R where R is alkyl, heterocycloalkyl, oraryl. The term “lower acyl” refers to where R is lower alkyl. The termC₁-C₄ acyl refers to where R is C₁-C₄.

The term “carboxy esters” refers to —C(O)OR where R is alkyl, aryl,aralkyl, cyclic alkyl, or heterocycloalkyl, all optionally substituted.

The term “carboxyl” refers to —C(O)OH.

The term “oxo” refers to ═O in an alkyl or heterocycloalkyl group.

The term “amino” refers to —NRR′ where R and R′ are independentlyselected from hydrogen, alkyl, aryl, aralkyl and heterocycloalkyl, allexcept H are optionally substituted; and R and R′ can form a cyclic ringsystem.

The term “-carboxylamido” refers to —CONR₂ where each R is independentlyhydrogen or alkyl.

The term “-sulphonylamido” or “-sulfonylamido” refers to —S(═O)₂NR₂where each R is independently hydrogen or alkyl.

The term “halogen” or “halo” refers to —F, —Cl, —Br and —I.

The term “alkylaminoalkylcarboxy” refers to the groupalkyl-NR-alk-C(O)—O— where “alk” is an alkylene group, and R is a H orlower alkyl.

The term “sulphonyl” or “sulfonyl” refers to —SO₂R, where R is H, alkyl,aryl, aralkyl, or heterocycloalkyl.

The term “sulphonate” or “sulfonate” refers to SO₂OR, where R is —H,alkyl, aryl, aralkyl, or heterocycloalkyl.

The term “alkenyl” refers to unsaturated groups which have 2 to 12atoms, 2 to 10 atom, or 2 to 8 atoms, and contain at least onecarbon-carbon double bond and includes straight-chain, branched-chainand cyclic groups. Alkenyl groups may be optionally substituted.Suitable alkenyl groups include allyl. “1-Alkenyl” refers to alkenylgroups where the double bond is between the first and second carbonatom. If the 1-alkenyl group is attached to another group, e.g. it is aW substituent attached to the cyclic phosphate, it is attached at thefirst carbon.

The term “alkynyl” refers to unsaturated groups which have 2 to 12atoms, 2 to 10 atoms, or 2 to 8 atoms, and contain at least onecarbon-carbon triple bond and includes straight-chain, branched-chainand cyclic groups. Alkynyl groups may be optionally substituted.Suitable alkynyl groups include ethynyl. “1-Alkynyl” refers to alkynylgroups where the triple bond is between the first and second carbonatom. If the 1-alkynyl group is attached to another group, e.g. it is aW substituent attached to the cyclic phosphate, it is attached at thefirst carbon.

The term “alkylene” refers to a divalent straight chain, branched chainor cyclic saturated aliphatic group. In one aspect the alkylene groupcontains up to and including 10 atoms. In another aspect, the alkylenechain contains up to and including 6 atoms. In a further aspect, thealkylene groups contains up to and including 4 atoms. The alkylene groupcan be either straight, branched or cyclic. The alkylene may beoptionally substituted with 1-3 substituents.

The term “acyloxy” refers to the ester group —O—C(O)R, where R is H,alkyl, alkenyl, alkynyl, aryl, aralkyl, or heterocycloalkyl.

The term “aminoalkyl-” refers to the group NR²-alk- wherein “alk” is analkylene group and R is selected from —H, alkyl, aryl, aralkyl, andheterocycloalkyl.

The term “alkylamino-” refers to the group alkyl-NR— wherein R is H oralkyl. “Lower alkylamino-” refers to groups where the alkyl is loweralkyl.

The term “alkylaminoalkyl-” refers to the group alkyl-NR-alk- whereineach “alk” is an independently selected alkylene, and R is H or loweralkyl. “Lower alkylaminoalkyl-” refers to groups where the alkyl and thealkylene group is lower alkyl and alkylene, respectively.

The term “arylaminoalkyl-” refers to the group aryl-NR-alk- wherein“alk” is an alkylene group and R is —H, alkyl, aryl, aralkyl, orheterocycloalkyl. In “lower arylaminoallyl-,” the alkylene group islower alkylene.

The term “alkylaminoaryl-” refers to the group alkyl-NR-aryl- wherein“aryl” is a divalent group and R is —H, alkyl, aralkyl, orheterocycloalkyl. In “lower alkylaminoaryl-,” the alkyl group is loweralkyl.

The term “alkoxyaryl-” refers to an aryl group substituted with analkyloxy group. In “lower alkyloxyaryl-”, the alkyl group is loweralkyl.

The term “aryloxyalkyl-” refers to an alkyl group substituted with anaryloxy group.

The term “aralkyloxyalkyl-” refers to the group aryl-alk-O-alk- wherein“alk” is an alkylene group. “Lower aralkyloxyalkyl-” refers to suchgroups where the alkylene groups are lower alkylene.

The term “alkoxy-” or “alkyloxy-” refers to the group alkyl-O—. In“lower alkoxy-,” each alkyl is lower alkyl.

The term “alkoxyalkyl-” or “alkyloxyalkyl-” refer to the groupalkyl-O-alk- wherein “alk” is an alkylene group. In “loweralkoxyalkyl-,” each alkyl and alkylene is lower alkyl and alkylene,respectively.

The terms “allylthio-” refers to the group alkyl-S—.

The term “alkylthioalkyl-” refers to the group alkyl-5-alk- wherein“alk” is an allkylene group. In “lower alkylthioalkyl-” each alkyl andalkylene is lower alkyl and alkylene, respectively.

The term “alkoxycarbonyloxy-” refers to alkyl-O—C(O)—O—.

The term “aryloxycarbonyloxy-” refers to aryl-O—C(O)—O—.

The term “alkylthiocarbonyloxy-” refers to alkyl-S—C(O)—O—.

The term “amido” refers to the NR₂ group next to an acyl or sulfonylgroup as in NR₂—C(O)—, RC(O)—NR¹—, NR₂—S(═O)₂— and RS(═O)₂—NR¹—, where Rand R¹ include —H, alkyl, aryl, aralkyl, and heterocycloalkyl. The term“carboxamido” refer to NR₂—C(O)— and RC(O)—NR¹—, where R and R¹ include—H, alkyl, aryl, aralkyl, and heterocycloalkyl. The term does notinclude urea, —NR—C(O)—NR—.

The terms “sulphonamido” or “sulfonamido” refer to NR₂—S(═O)₂— andRS(═O)₂—NR¹—, where R and R¹ include —H, alkyl, aryl, aralkyl, andheterocycloalkyl. The term does not include sulfonylurea,—NR—S(═O)₂—NR—.

The term “carboxamidoalkylaryl” and “carboxamidoaryl” refers to anaryl-alk-NR¹—C(O), and ar-NR¹—C(O)-alk-, respectively where “ar” isaryl, “alk” is alkylene, R¹ and R include H, alkyl, aryl, aralkyl, andheterocycloalkyl.

The term “sulfonamidoalkylaryl” and “sulfonamidoaryl” refers to anaryl-alk-NR¹—S(═O)₂—, and ar-NR¹—S(═O)₂—, respectively where “ar” isaryl, “alk” is alkylene, R¹ and R include —H, alkyl, aryl, aralkyl, andheterocycloalkyl.

The term “hydroxyalkyl” refers to an alkyl group substituted with one—OH.

The term “haloalkyl” refers to an alkyl group substituted with onehalogen.

The term “cyano” refers to —CN═.

The term “nitro” refers to —NO₂.

The term “acylalkyl” refers to an allyl-C(O)-alk-, where “alk” isalkylene.

The term “aminocarboxamidoalkyl-” refers to the group NR₂—C(O)—N(R)-alk-wherein R is an alkyl group or H and “alk” is an alkylene group. “Loweraminocarboxamidoalkyl-” refers to such groups wherein “alk” is loweralkylene.

The term “heteroarylalkyl” refers to an alkylene group substituted witha heteroaryl group.

The term “perhalo” refers to groups wherein every C—H bond has beenreplaced with a C-halo bond on an aliphatic or aryl group. Suitableperhaloalkyl groups include —CF₃ and —CFCl₂.

The term “purine” refers to nitrogenous bicyclic heterocycles. The term“pyrimidine” refers to nitrogenous monocyclic heterocycles. The term“purine” or “pyrimidine” base includes, but is not limited to, adenine,N⁶-alkylpurines, N⁶-acylpurines (wherein acyl is C(O)(alkyl, aryl,alkylaryl, or arylalkyl), N⁶-benzylpurine, N⁶-halopurine,N⁶-vinylpurine, N⁶-acetylenic purine, N⁶-acyl purine, N⁶-hydroxyalkylpurine, N⁶-thioalkyl purine, N²-alkylpurines, N²-alkyl-6-thiopurines,thymine, cytosine, 5-fluorocytosine, 5-methylcytosine, 6-azapyrimidine,including 6-azacytosine, 2- and/or 4 mercaptopyrmidine, uracil,5-halouracil, including 5-fluorouracil, C⁵-alkylpyrimidines,C⁵-benzylpyrimidines, C⁵-halopyrimidines, C⁵-vinylpyrimidine,C⁵-acetylenic pyrimidine, C⁵-acyl pyrimidine, C⁵-hydoxyalkyl purine,C⁵-amidopyrimidine, C⁵-cyanopyrimidine, C⁵-mtropyrimidine,C⁵-aminopyrimidine-, N²-alkylpurines, N²-alkyl-6-thiopurines,5-azacytidinyl, 5-azauracilyl, triazolopyridinyl, imidazolopyridinyl,pyrrolopyrimidinyl, and pyrazolopyrimidinyl. Purine bases include, butare not limited to, guanine, adenine, hypoxanthine, 2,6-diaminopurine,and 6-chloropurine. Functional oxygen and nitrogen groups on the basecan be protected as necessary or desired. Suitable protecting groups arewell known to those skilled in the art, and include trimethylsilyl,dimethylhexylsilyl, t-butyldimethylsilyl, and t-butyldiphenylsilyl,trityl, alkyl groups, and acyl groups such as acetyl and propionyl,methanesulfonyl, and p-toluenesulfonyl.

The phrase “therapeutically effective amount” means an amount of acompound or a combination of compounds that ameliorates, attenuates oreliminates one or more of the symptoms of a particular disease orcondition or prevents, modifies, or delays the onset of one or more ofthe symptoms of a particular disease or condition.

The term “pharmaceutically acceptable salt” includes salts of a compoundof the present invention including compounds of Formulae I-XVI and itsprodrugs derived from the combination of a compound of this inventionand an organic or inorganic acid or base. Suitable acids include aceticacid, adipic acid, benzenesulfonic acid,(+)-7,7-dimethyl-2-oxobicyclo[2.2.1]heptane-1-methanesulfonic acid,citric acid, 1,2-ethanedisulfonic acid, dodecyl sulfonic acid, fumaricacid, glucoheptonic acid, gluconic acid, glucuronic acid, hippuric acid,hydrochloride hemiethanolic acid, HBr, HCl, HI, 2-hydroxyethanesulfonicacid, lactic acid, lactobionic acid, maleic acid, methanesulfonic acid,methylbromide acid, methyl sulfuric acid, 2-naphthalenesulfonic acid,nitric acid, oleic acid, 4,4′-methylenebis[3-hydroxy-2-naphthalenecarboxylic acid], phosphoric acid,polygalacturonic acid, stearic acid, succinic acid, sulfuric acid,sulfosalicylic acid, tannic acid, tartaric acid, terphthalic acid, andp-toluenesulfonic acid.

The term “naturally-occurring L-amino acid” refers to those amino acidsroutinely found as components of proteinaceous molecules in nature,including alanine, valine, leucine, isoleucine, proline, phenylalanine,tryptophan, methionine, glycine, serine, threonine, cysteine, tyrosine,asparagine, glutamine, aspartic acid, glutamic acid, lysine, arginineand histidine. In one aspect, this term is intended to encompass L-aminoacids having only the amine and carboxylic acid as charged functionalgroups, i.e., alanine, valine, leucine, isoleucine, proline,phenylalanine, tryptophan, methionine, glycine, serine, threonine,cysteine and tyrosine. In another aspect they are alanine, valine,leucine, isoleucine, proline, phenylalanine, and glycine. In a furtheraspect, it is valine.

The term “ester of an L-amino acid” refers to ester formed by couplingof a hydroxyl group of the compound with a carboxylic acid of naturallyoccurring L-amino acid.

The term “patient” refers to an animal being treated including a mammal,such as a dog, a cat, a cow, a horse, a sheep, and a human. Anotheraspect includes a mammal, both male and female.

The term “prodrug” as used herein refers to any compound that whenadministered to a biological system generates a biologically activecompound as a result of spontaneous chemical reaction(s), enzymecatalyzed chemical reaction(s), and/or metabolic chemical reaction(s),or a combination of each. Standard prodrugs are formed using groupsattached to a functionality, e.g. HO—, HS—, HOOC—, R₂N—, associated withthe drug, that cleave in vivo. Standard prodrugs include but are notlimited to carboxylate esters where the group is alkyl, aryl, aralkyl,acyloxyalkyl, alkoxycarbonyloxyalkyl as well as esters of hydroxyl,thiol and amines where the group attached is an acyl group, analkoxycarbonyl, aminocarbonyl, phosphate or sulfate.

For example, phosphonate or monophosphate prodrugs are compounds thatbreakdown chemically or enzymatically to a phosphonic acid ormonophosphate or phosphinic acid group or a monoester thereof in vivo.As employed herein the term includes, but is not limited to, thefollowing groups and combinations of these groups:

Acyloxyalkyl esters which are well described in the literature (Farquharet al., J. Pharm. Sci. 72:324-325 (1983)).

Other acyloxyalkyl esters are possible in which a cyclic alkyl ring isformed. These esters have been shown to generate phosphorus-containingnucleotides inside cells through a postulated sequence of reactionsbeginning with deesterification and followed by a series of eliminationreactions (e.g., Freed et al., Biochem. Pharm, 38:3193-3198 (1989)).

Another class of these double esters known as alkyloxycarbonyloxymethylesters, as shown in formula A, where R′ is alkoxy, aryloxy, alkylthio,arylthio, alkylamino, and arylamino; R′, and R″ are independently —H,alkyl, aryl, alkylaryl, and heterocycloalkyl have been studied in thearea of β-lactam antibiotics (Nishimura et al., J. Antibiotics40(1):81-90 (1987); for a review see Ferres, H., Drugs of Today, 19:499(1983)). More recently Cathy, M. S. et al. (Abstract from AAPS WesternRegional Meeting, April, 1997) showed that thesealkyloxycarbonyloxymethyl ester prodrugs on(9-[(R)-2-phosphonomethoxy)propyl]adenine (PMPA) are bioavailable up to30% in dogs.

wherein R, R′, and R″ are independently H, alkyl, aryl, alkylaryl, andalicyclic (see, e.g., International Publ. Nos. WO 90/08155 and WO90/10636).

Other acyloxyalkyl esters are possible in which a cyclic alkyl ring isformed such as shown in Formula B. These esters have been shown togenerate phosphorus-containing nucleotides inside cells through apostulated sequence of reactions beginning with deesterification andfollowed by a series of elimination reactions (see, e.g., Freed et al.,Biochem. Pharm. 38:3193-3198 (1989)).

wherein R is —H, alkyl, aryl, alkylaryl, alkoxy, aryloxy, alkylthio,arylthio, alkylamino, arylamino, or cycloalkyl.

Aryl esters have also been used as phosphorus prodrugs (e.g., DeLambertet al., J. Med. Chem. 37(7):498-511 (1994); Serafinowska et al., J. Med.Chem. 38(8):1372-1379 (1995)). Phenyl as well as mono andpoly-substituted phenyl proesters have generated the parent phosphonicacid in studies conducted in animals and in man (Formula C). Anotherapproach has been described where Y is a carboxylic ester ortho to thephosphate (Khamnei et al., J. Med. Chem. 39:4109-4115 (1996)).

wherein Y is —H, alkyl, aryl, alkylaryl, alkoxy, acyloxy, halogen,amino, alkoxycarbonyl, hydroxy, cyano, and heterocycloalkyl.

Benzyl esters have also been reported to generate the parent phosphonicacid. In some cases, using substituents at the para-position canaccelerate the hydrolysis. Benzyl analogs with 4-acyloxy or 4-alkyloxygroup (Formula D below, wherein X≡—H, OR or O(CO)R or O(CO)OR) cangenerate the 4-hydroxy compound more readily through the action ofenzymes, e.g., oxidases, esterases, etc. Examples of this class ofprodrugs are described in Mitchell et al., J. Chem. Soc. Perkin Trans.12345 (1992); International Publ. No. WO 91/19721.

wherein X and Y are independently —H, alkyl, aryl, alkylaryl, alkoxy,acyloxy, hydroxy, cyano, nitro, perhaloalkyl, halo, or alkyloxycarbonyl;and R′ and R″ are independently —H, alkyl, aryl, alkylaryl, halogen, andcyclic alkyl.

Thio-containing phosphonate proesters may also be useful in the deliveryof drugs to hepatocytes. These proesters contain a protected thioethylmoiety as shown in Formula E. One or more of the oxygens of thephosphonate can be esterified. Since the mechanism that results inde-esterification requires the generation of a free thiolate, a varietyof thiol protecting groups are possible. For example, the disulfide isreduced by a reductase-mediated process (Puech et al., Antiviral Res.22:155-174 (1993)). Thioesters will also generate free thiolates afteresterase-mediated hydrolysis Benzaria, et al., J. Med. Chem.39(25):4958-4965 (1996)). Cyclic analogs are also possible and wereshown to liberate phosphonate in isolated rat hepatocytes. The cyclicdisulfide shown below has not been previously described and is novel.

wherein Z is alkylcarbonyl, alkoxycarbonyl, arylcarbonyl,aryloxycarbonyl, or alkylthio.

Other examples of suitable prodrugs include proester classes exemplifiedby Biller and Magnin U.S. Pat. No. 5,157,027); Serafinowska et al., J.Med. Chem. 38(8):1372-1379 (1995); Starrett et al., J. Med. Chem.37:1857 (1994); Martin et al., J. Pharm. Sci. 76:180 (1987); Alexanderet al., Collect. Czech. Chem. Commun. 59:1853 (1994); and EP 0 632 048A1. Some of the structural classes described are optionally substituted,including fused lactones attached at the omega position (Formulae E-1and E-2) and optionally substituted 2-oxo-1,3-dioxolenes attachedthrough a methylene to the phosphorus oxygen (Formula E-3) such as:

wherein R is —H, alkyl, cycloalkyl, or heterocycloalkyl; and wherein Yis —H, alkyl, aryl, alkylaryl, cyano, alkoxy, acyloxy, halogen, amino,heterocycloalkyl, and alkoxycarbonyl.

The prodrugs of Formula E-3 are an example of “optionally substitutedheterocycloalkyl where the cyclic moiety contains a carbonate orthiocarbonate.”

Propyl phosphonate proesters can also be used to deliver drugs intohepatocytes. These proesters may contain a hydroxyl and hydroxyl groupderivatives at the 3-position of the propyl group as shown in Formula F.The R and X groups can form a cyclic ring system as shown in Formula F.One or more of the oxygens of the phosphonate can be esterified.

wherein R is alkyl, aryl, heteroaryl; X is hydrogen, alkylcarbonyloxy,alkyloxycarbonyloxy; and Y is alkyl, aryl, heteroaryl, alkoxy,alkylamino, alkylthio, halogen, hydrogen, hydroxy, acyloxy, amino.

Phosphoramidate derivatives have been explored as phosphate prodrugs(e.g., McGuigan et al., J. Med. Chem. 42:393 (1999) and references citedtherein) as shown in Formula G and H.

Cyclic phosphoramidates have also been studied as phosphonate prodrugsbecause of their speculated higher stability compared to non-cyclicphosphoramidates (e.g., Starrett et al., J. Med. Chem. 37:1857 (1994)).

Another type of phosphoramidate prodrug was reported as the combinationof S-acyl-2-thioethyl ester and phosphoramidate (Egron et al.,Nucleosides Nucleotides 18:981 (1999)) as shown in Formula 3:

Other prodrugs are possible based on literature reports such assubstituted ethyls, for example, bis(trichloroethyl)esters as disclosedby McGuigan, et al., Bioorg Med. Chem. Lett. 3:1207-1210 (1993), and thephenyl and benzyl combined nucleotide esters reported by Meier, C. etal., Bioorg. Med. Chem. Lett. 7:99-104 (1997).

The groups illustrated are exemplary, not exhaustive, and one skilled inthe art could prepare other known varieties of prodrugs. Such prodrugsof the compounds of Formula I fall within this scope. Prodrugs mustundergo some form of a chemical transformation to produce the compoundthat is biologically active or is a precursor of the biologically activecompound. In some cases, the prodrug is biologically active, usuallyless than the drug itself, and serves to improve drug efficacy or safetythrough improved oral bioavailability, pharmacodynamic half-life, etc.Prodrug forms of compounds may be utilized, for example, to improvebioavailability, improve subject acceptability such as by masking orreducing unpleasant characteristics such as bitter taste orgastrointestinal irritability, alter solubility such as for intravenoususe, provide for prolonged or sustained release or delivery, improveease of formulation, or provide site-specific delivery of the compound.Prodrugs are described in The Organic Chemistry of Drug Design and DrugAction, by Richard B. Silverman, Academic Press, San Diego (1992);Chapter 8: “Prodrugs and Drug delivery Systems,” pp. 352-401; Design ofProdrugs, edited by H. Bundgaard, Elsevier Science, Amsterdam (1985);Design of Biopharmaceutical Properties through Prodrugs and Analogs, Ed.by E. B. Roche, American Pharmaceutical Association, Washington (1977);and Drug Delivery Systems, ed. by R. L. Juliano, Oxford Univ. Press,Oxford (1980).

In the case of bases, “prodrugs” are preferred at the 6-position ofpurine analogs. Such substitution may include H, halogen, amino, acetoxyor azido or alkyl carbamoyl groups. Hydrogen substituted prodrugs at the6-position of guanosine analogs undergo oxidation in vivo by aldehydeoxidase or xanthine oxidase to give the required functionality (Rashidiet al., Drug Metab. Dispos. 25:805 (1997)). While esterases unmaskacetoxy groups, amine and halogen substituents are known to besubstrates for deaminases. 6-Azido substituted compounds are also knownto give the corresponding amino derivatives by the action of reductases(Koudriakova, et al., J. Med Chem. 39:4676 (1996)).

The structure

has a plane of symmetry running through the phosphorus-oxygen doublebond when V=W and V and W are either both pointing up or both pointingdown.

The term “cyclic phosphate ester of 1,3-propanediol,”, “cyclic phosphatediester of 1,3-propanediol,” “2 oxo 2λ⁵ [1,3,2] dioxaphosphorinane,”“2-oxo-[1,3,2]-dioxaphosphorinane,” or “dioxaphosphorinane” refers tothe following:

The phrase “V and Z are connected together via an additional 3-5 atomsto form a cyclic group, optionally containing one heteroatom, that isfused to an aryl group attached at the beta and gamma position to the 0attached to the phosphorus” includes the following:

As shown above V and Z are connected together via 4 additional atoms.

The phrase “W and W′ are connected together via an additional 2-5 atomsto form a cyclic group, optionally containing 0-2 heteroatoms, and Vmust be aryl, substituted aryl, heteroaryl, or substituted heteroaryl”includes the following:

As shown above W and W′ are connected together via an additional 2atoms. The structure above has V=aryl, and a spiro-fused cyclopropylgroup for W and W′.

The term “cyclic phosphate” refers to

The carbon attached to V must have a C—H bond. The carbon attached to Zmust also have a C—H bond.

The term “cis” stereochemistry refers to the spatial relationship of theV group and the substituent attached to the phosphorus atom via anexocyclic single bond on the six membered 2-oxo-phosphorinane ring. Thestructures K and L below show two possible cis-isomers of 2- and4-substituted 2-oxo-phosphorinane. Structure K shows cis-isomer of (2S,4R)-configuration whereas structure L shows cis-isomer of(2R,4S)-configuration.

The term “trans” stereochemistry refers to the spatial relationship ofthe V group and the substituent attached to the phosphorus atom via anexocyclic single bond on the six membered 2-oxo-phosphorinane ring. Thestructures M and N below show two possible trans-isomers of 2- and4-substituted 2-oxo-phosphorinane. Structure M shows trains-isomer of(2S, 4S)— configuration whereas structure N shows trans-isomer of(2R,4R)-configuration.

The term “percent enantiomeric excess (% ee)” refers to optical purity.It is obtained by using the following formula:

${\frac{\lbrack R\rbrack - \lbrack S\rbrack}{\lbrack R\rbrack + \lbrack S\rbrack} \times 100} = {{\% \mspace{14mu} R} - {\% \mspace{14mu} S}}$

where [R] is the amount of the R isomer and [S] is the amount of the Sisomer. This formula provides the % ee when R is the dominant isomer.

The term “enantioenriched” or “enantiomerically enriched” refers to asample of a chiral compound that consists of more of one enantiomer thanthe other. The extent to which a sample is enantiomerically enriched isquantitated by the enantiomeric ratio or the enantiomeric excess.

The term “liver” refers to liver organ.

The term “enhancing” refers to increasing or improving a specificproperty.

The term “liver specificity” refers to the ratio:

$\frac{\left\lbrack {{drug}\mspace{14mu} {or}\mspace{14mu} a\mspace{14mu} {drug}\mspace{14mu} {metabolite}\mspace{14mu} {in}\mspace{14mu} {liver}\mspace{14mu} {tissue}} \right\rbrack}{\left\lbrack {{drug}\mspace{14mu} {or}\mspace{14mu} a\mspace{14mu} {drug}\mspace{14mu} {metabolite}\mspace{14mu} {in}\mspace{14mu} {blood}\mspace{14mu} {or}\mspace{14mu} {another}\mspace{14mu} {tissue}} \right\rbrack}$

as measured in animals treated with the drug or a prodrug. The ratio canbe determined by measuring tissue levels at a specific time or mayrepresent an AUC based on values measured at three or more time points.

The term “increased or enhanced liver specificity” refers to an increasein the liver specificity ratio in animals treated with the prodrugrelative to animals treated with the parent drug.

The term “enhanced oral bioavailability” refers to an increase of atleast 50% of the absorption of the dose of the parent drug. In anadditional aspect the increase in oral bioavailability of the prodrug(compared to the parent drug) is at least 100%, that is a doubling ofthe absorption. Measurement of oral bioavailability usually refers tomeasurements of the prodrug, drug, or drug metabolite in blood, plasma,tissues, or urine following oral administration compared to measurementsfollowing parenteral administration.

The term “therapeutic index” refers to the ratio of the dose of a drugor prodrug that produces a therapeutically beneficial response relativeto the dose that produces an undesired response such as death, anelevation of markers that are indicative of toxicity, and/orpharmacological side effects.

The term “sustained delivery” refers to an increase in the period inwhich there is a prolongation of therapeutically-effective drug levelsdue to the presence of the prodrug.

The term “bypassing drug resistance” refers to the loss or partial lossof therapeutic effectiveness of a drug (drug resistance) due to changesin the biochemical pathways and cellular activities important forproducing and maintaining the biological activity of the drug and theability of an agent to bypass this resistance through the use ofalternative pathways or the failure of the agent to induce changes thattend to resistance.

The terms “treating” or “treatment” of a disease includes inhibiting thedisease (slowing or arresting its development), providing relief fromthe symptoms or side-effects of the disease (including palliativetreatment), and relieving the disease (causing regression of thedisease).

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to compounds of Formula I, and isomers,hydrates, solvates, prodrugs, co-crystals, and pharmaceuticallyacceptable salts thereof:

wherein:

X′ is O, S, S—O, or NR²⁰, wherein R²⁰ is H or optionally substitutedalkyl, aryl, arylalkyl, C₃₋₆ cycloalkyl, OH, OR^(20′), or O(C═O)R^(20′),wherein R^(20′) is H, lower alkyl or C₃₋₆ cycloalkyl;

Y is —O—, —S—, —N—, —C(R^(20′))—, or —CH₂—;

R¹⁹ is H or optionally substituted C₁₋₄ alkyl, C₂₋₄ alkenyl, or C₂₋₄alkynyl, —OH, —O-lower alkyl, halogen, CN, or —C═CR²¹R²², wherein R²¹and R²² are independently H or lower alkyl;

or R¹⁹ is absent; or R¹⁹ is joined together with R¹⁷ to form—(CH₂)_(p)—, —O—(CH₂)_(p)—, wherein p is 0 to 4;

R¹⁸ is independently H, C₁₋₄ alkyl, C₂₋₄ alkenyl, or C₂₋₄ alkynyl;wherein said C₁₋₄ alkyl is optionally substituted with amino, hydroxy,or 1 to 3 fluorine atoms, C₁₋₄ alkylamino, dialkylamino, C₃₋₆cycloalkylamino, halogen, or alkoxy;

R¹⁷ is H, halogen, alkyl optionally substituted with 1 to 3 fluorineatoms, C₁₋₁₀ alkoxy optionally substituted with C₁₋₃ alkoxy or 1 to 3fluorine atoms, C₂₋₆ alkenyloxy, C₁₋₄ alylthio, C₁₋₈ alkylcarbonyloxy,aryloxycarbonyl, azido, amino, alkylamino, or dialkylamino;

R¹⁶ and R¹⁵ are independently H, C₁₋₄ alkyl, C₂₋₄ alkenyl, or C₂₋₄alkynyl; wherein said C₁₋₄ alkyl is optionally substituted with amino,hydroxy, or 1 to 3 fluorine atoms, and said C₂₋₄ alkenyl and C₂₋₄alkynyl are each optionally substituted with one or more of C₁₋₃ alkoxy,carboxy, C₂₋₆ alkenyloxy, C₁₋₄ alkylthio, C₁₋₈ alkylcarbonyloxy,aryloxycarbonyl, azido, amino, alkylamino, or dialkylamino;

B is a purine or pyrimidine base or an analogue or derivative thereof;

Z₁ is —CH(R²³)—OH, —O—, —CH(R²³)—O—, C₁₋₄ cycloalkyl, —OC(R²³)₂PO₃H₂,—CH₂C(R²³)₂PO₃H₂, C₂₋₄ alkenyl, C₂₋₄ alkynyl, C₁₋₄ cycloalkylene, C₂₋₄alkenylene, or C₂₋₄ alkynylene; wherein R²³ is H, F, methyl, ethyl,hydroxymethyl, fluoromethyl, —CH₂N₃, —CH₂—NR²¹R²², —CH₂—, or —CH₂—NH₂;and

Z″ is absent, or Z″ is R²⁴(C═O)—, R²⁴—O—(C═O)—, or an ester of anL-amino acid such as an L-valine ester R²⁴CH(NH₂)(C═O)—, wherein R²⁴ isoptionally substituted C₁₋₆ alkyl, cycloalkyl, aryl, or aralkyl; or Z″is

wherein:

V, W, and W′ are independently H, optionally substituted allyl,optionally substituted aralkyl, cycloalkyl, heterocycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, optionallysubstituted 1-alkenyl, or optionally substituted 1-alkynyl; and

Z is —CHR^(z)OH, —CH^(z)OC(O)R^(y), —CHR^(z)OC(S)R^(y),—CHR^(z)OC(S)OR^(y), —CHR^(z)OC(O)SR^(y), —CHR^(z)OCO₂R^(y), —OR^(z),—SR^(z), —CHR^(z)N3, —CH₂aryl, —CH(aryl)OH, —CH(CH═CR^(z) ₂)OH,—CH(C≡CR^(z))OH, —R^(z), —NR^(z2), —OCOR^(y), —OCO₂R^(y), —SCOR^(y),—SCO₂R^(y), —NHCOR^(z), —NHCO₂R^(y), —CH₂NHaryl, —(CH₂)_(q)—OR^(z), and—(CH₂)_(q)—SR^(z), halogen, —CN, —COR^(y), —CONR^(z) ₂, —CO₂R^(y),—SO₂R^(y), or —SO₂NR^(z) ₂, wherein q is 2 or 3, R^(z) is R^(y) or —H,and R^(y) is alkyl, aryl, cycloalkyl, heterocycloalkyl, or aralkyl; or

Z″ is P(O)Y′R¹¹Y″R¹¹, wherein each R¹¹ is independently H or C₁₋₄ alkyl;Y and Y′ are each independently selected from the group consisting of—O—, and —NR^(v)—; and

when Y′ and Y″ are both —O—, R¹¹ attached to —O— is independentlyselected from the group consisting of optionally substituted aryl,optionally substituted CH₂-heterocycloakyl wherein the cyclic moietycontains a carbonate or thiocarbonate, optionally substituted-alkylaryl, —C(R^(z))₂OC(O)NR^(z) ₂, —NR^(z)—C(O)—R^(y),—(R^(z))₂—OC(O)R^(y), —C(z)₂—O—C(O)OR^(y), —C(z)₂OC(O)SR^(y),-alkyl-S—C(O)R^(y), -alkyl-S—S-alkylhydroxy, and-alkyl-S—S—S-alkylhydroxy; or

when Y′ and Y″ are both —N^(v)—, then R¹¹ attached to —NR^(v)— isindependently selected from the group consisting of —H,—[C(R^(z))₂]_(q)—COOR^(y), —C(R^(x))₂COOR^(y),—[C(R^(z))₂]_(q)—C(O)SR^(y) and -cycloalkylene-COOR^(y); or

when Y′ is —O— and Y″ is NR^(v), then R¹¹ attached to —O— isindependently selected from the group consisting of optionallysubstituted aryl, optionally substituted CH₂-heterocycloakyl wherein thecyclic moiety contains a carbonate or thiocarbonate, optionallysubstituted -alkylaryl, —C(R^(z))₂OC(O)NR^(z) ₂, —NR^(z)—C(O)—R^(y),—C(R^(z))₂—OC(O)R^(y), —C(R^(z))₂—O—C(O)OR^(y), —C(z)₂OC(O)SR^(y),-alkyl-S—C(O)R^(y), -alkyl-S—S-alkylhydroxy, and-alkyl-S—S—S-alkylhydroxy; and R¹¹ attached to —NR^(v)— is independentlyselected from the group consisting of —H, —[C(R^(z))₂]_(q)—COOR^(y),—C(R^(x))₂COOR^(y), —[C(R^(z))₂]_(q)—C(O)SR^(y), and-cycloalkylene-COOR^(y); or

when Y′ and Y″ are independently selected from —O— and —NR^(v)—, thenR¹¹ and R¹¹ together form a cyclic group comprising -alkyl-S—S-alkyl-;

wherein q is an integer 2 or 3;

each R^(z) is selected from the group consisting of R^(y) and —H;

each R^(y) is selected from the group consisting of alkyl, aryl,heterocycloalkyl, and aralkyl;

each R^(x) is independently selected from the group consisting of —H,and alkyl, or together R^(x) and R^(x) form a cycloalkyl group; and

each R^(v) is selected from the group consisting of —H, lower alkyl,acyloxyalkyl, alkoxycarbonyloxyalkyl, and lower acyl.

In some aspects of the present invention, the following provisos apply:

(a) V, Z, W, W′ are not all —H; (b) when Z is —R^(z), then at least oneof V, W, and W′ is not —H, alkyl, aralkyl, cycloalkyl, orheterocycloalkyl; c) when Z′ is —CH₂OH and R¹⁷ is H, then one of R¹⁵,R¹⁶, R¹⁷ and R¹⁸ is other than H; and d) when Z′ is —CH₂O—, Z″ is—(C═O)R²⁴, and R¹⁷ is H, then one of R¹⁵, R¹⁶, R¹⁷ and R¹⁸ is other thanH.

In some aspects of the present invention, compounds of Formula I arethose in which X′ is O or S. For example, in some aspects, compounds ofFormula I are those in which X′ is O. In other aspects, compounds ofFormula I are those in which X′ is S.

In some aspects, compounds of Formula I are those in which X′ is NR²⁰.In these aspects, suitable values of R²⁰ include H, C₁₋₁₀ alkyl, C₆₋₁₀aryl, C₆₋₁₀ aryl(C₁₋₆)alkyl, or C₃₋₆ cycloalkyl. In other aspects,suitable values of R²⁰ include OH, OR^(20′), or O(C═O)R^(20′), whereinR^(20′) is H, C₁₋₆ alkyl or C₃₋₆ cycloalkyl

In some aspects, compounds of Formula I are those in which Y is —O—,—S—, —N—, or —CH₂—. In other aspects, Y is —C(R^(20′))—. In yet otheraspects of the invention, Y is —O—.

In some aspects, R¹⁹ is absent in compounds of Formula I. In otheraspects, R¹⁹ is present and is H, —OH, —O-lower alkyl, e.g., —OCH₃, orR¹⁹ is optionally substituted C₁₋₄ alkyl, e.g., methyl. In yet otheraspects, R¹⁹ is joined together with R¹⁷ to form —O—(CH₂)_(p), wherein pis 2 or 3.

In some aspects, R¹⁸ is H, C₁₋₄ alkyl, wherein said C₁₋₄ alkyl isoptionally substituted with amino, hydroxy, or 1 to 3 fluorine atoms. Inother aspects, R¹⁸ is H or C14 alkyl, e.g., methyl or ethyl.

In some aspects, R¹⁸, R¹⁷, R¹⁶, and R¹⁵ are independently H or loweralkyl, e.g., C₁₋₆ alkyl, such as methyl, ethyl, or propyl. For example,in some aspects, R¹⁶ is —CH₃.

The variable B depicted in Formula I above represents a purine orpyrimidine base or analogue or derivative thereof B will be preferablylinked to the ribose ring of Formula I at the 9- or 1-position,respectively, of the purine or pyrimidine base B. By “purine orpyrimidine base or analogue or derivative thereof” is meant a purine orpyrimidine base found in native nucleosides, or an analogue thereof,which mimics such bases in that their structures (the kinds of atoms andtheir arrangement) are similar to the native bases but may eitherpossess additional or lack certain of the functional properties of thenative bases. Such analogues include those derived by replacement of aCH moiety by a nitrogen atom (for example, 5-azapyrimidines such as5-azacytosine) or vice versa (for example, 7-deazapurines such as7-deazadenine or 7-deazaguanine) or both (e.g., 7-deaza, 8-azapurines).By derivatives of such bases or analogues are meant those compoundswherein ring substituents are either incorporated, removed, or modifiedby conventional substituents known in the art, e.g., halogen, hydroxyl,amino, and C₁₋₆ alkyl. Such purine or pyrimidine bases, analogues, andderivatives will be well known to those skilled in the art.

Thus, in some aspects of the present invention, B is selected from:

wherein:

A, D, E, J, and G are each independently selected from the groupconsisting of C and N;

L is selected from O or S;

M is selected from the group consisting of O, S, and Se;

X₁ is absent, or X₁ is selected from the group consisting of H, —OH,—SH, —NH₂, —CO, —COOR¹¹, —CONH₂, —CSNH₂, alkylamino, dialkylamino,cycloalkylamino, halogen, alkyl, alkenyl, alkynyl, aryl, alkaryl,cycloalkyl, acyl, alkoxy, CF₃, and —NHCOR_(X1), wherein R_(X1) is H,lower alkyl, or lower alkoxy, and wherein R¹¹ is H or C₁₋₄ alkyl;

X₂ is absent, or X₂ is independently selected from the group consistingof H, alkenyl, alkynyl, aryl, alkaryl, cycloalkyl, acyl, and C₁-C₆alkyl;

X₃, X₄ and X₆ are each independently absent, or X₃, X₄ and X₆ are eachindependently selected from the group consisting of H, alkenyl, alkynyl,aryl, alkaryl, cycloalkyl, acyl, OH, SH, NH₂, CF₃, alkyl, amino,halogen, alkylamino, cycloalkylamino, and dialkylamino; and

X₅ is absent, or X₅ is selected from the group consisting of H, —CN,—NO₂, -alkyl, alkenyl, alkynyl, aryl, alkaryl, cycloalkyl, acyl,—NHCONH₂, —CONR¹¹R^(11′), —CSNR¹¹R^(11′), —COOR^(11′), —C(═NH)NH₂,-hydroxy, —C₁₋₃ alkoxy, -amino, -alkylamino, -dialkylamino, halogen,-(1,3-oxazol-2-yl), -(1,3-thiazol-2-yl), and -(imidazol-2-yl); whereinalkyl is unsubstituted or substituted with one to three groupsindependently selected from the group consisting of halogen, amino,hydroxy, carboxy, and C₁₋₃ alkoxy; and wherein R¹¹ and R^(11′) areindependently H or C₁₋₄ alkyl.

In one aspect of the present invention, B is selected from:

wherein R²⁵ is independently selected from the group consisting of H andNH₂; and R²⁶ is selected from the group consisting of NH₂, NHCH₃,N(CH₃)₂, OCH₃, SCH₃, OH, Cl, Br, SH, cyclopropyl amino, cyclobutylamino, and cyclopentyl amino.

In other aspects, B is selected from:

In another aspect of the present invention, B is selected from thefollowing:

wherein:

each R₄ and R₅ is independently H, acyl, C₁-C₆ alkyl, alkenyl, alkynylor cycloalkyl;

W₁, W₂, W₃ and W₄ are each independently N, CH, CF, Cl, CBr, CCl, CCN,CCH₃, CCF₃, CCH₂CH₃, CC(O)NH₂, CC(O)N(R₄)₂, CC(O)OH, CC(O)OR₄ or CT₃;wherein T₃ is as defined below;

W₅ is O, S, NH or NR₄;

T₂ is H, optionally substituted alkyl (such as, e.g., CH₃, CF₃, C(Y₃)₃,2-Br-ethyl, CH₂F, CH₂C₁, CF₂CF₃, C(Y₃)₂C(Y₃)₃, or CH₂OH), optionallysubstituted alkenyl, optionally substituted alkynyl, COOH, COOR₄,COO-alkyl, COO-aryl, CO-alkoxyalkyl, CONH₂, CONHR₄, CON(R₄)₂, chloro,bromo, fluoro, iodo, CN, N₃, OH, OR₄, NH₂, NHR₄, NR₄R₅, SH or SR₅,wherein Y₃ is as defined below;

T₃ is optionally substituted alkyl (including lower alkyl, such as,e.g., CH₃, CH₂CN, CH₂N₃, CH₂NH₂, CH₂NHCH₃, CH₂N(CH₃)₂, CH₂₀H),halogenated alkyl (including halogenated lower alkyl such as, e.g., CF₃,C(Y₃)₃, 2-Br-ethyl, CH₂F, CH₂Cl, CH₂CF₃, CF₂CF₃, C(Y₃)₂C(Y₃)₃),optionally substituted alkenyl, haloalkenyl, optionally substitutedalkynyl, haloalkynyl, Br-vinyl, N₃, CN, —C(O)OH, —C(O)OR₄, —C(O)O(loweralkyl), —C(O)NH₂, —CONHR₄, —C(O)NH(lower alkyl), —C(O)N(R₄)₂,—C(O)N(lower alkyl)₂, OH, OR₄, —O(acyl), —O(lower acyl), —O(alkyl),—O(lower alkyl), —O(alkenyl), —O(alkynyl), —O(aralkyl), —O(cycloalkyl),—S(acyl), —S(lower acyl), —S(alkyl), —S(lower alkyl), —S(alkenyl),—S(alkynyl), —S(aralkyl), —S(cycloalkyl), chloro, bromo, iodo, fluoro,NH₂, NH₄, NR₄R₅, —NH(lower alkyl), —NH(acyl), —N(lower alkyl)₂,—NH(alkenyl), —NH(alkynyl), —NH(aralkyl), —O(cycloalkyl), or —N(acyl)₂,wherein Y₃ is as defined below;

Y₁, is H, Br, Cl, I, F, CN, OH, OR₄, NH₂, NHR₄, NR₄R₅, SH or SR₄,wherein R₄ and R₅ are as defined below;

Y₂ is O, S, NH or NR₄, wherein R₄ is as defined below;

Y₃ is H, Br, Cl, I, F;

Y₄ is H, optionally substituted lower alkyl, cycloalkyl, alkenyl,alkynyl, CH₂OH, CH₂NH₂, CH₂NHCH₃, CH₂N(CH₃)₂, CH₂F, CH₂Cl, CH₂N₂, CH₂CN,CH₂CF₃, CF₃, CF₂CF₃, CH₂CO₂R, (CH₂)_(m)COOH, (CH₂)_(m)COOR,(CH₂)_(m)CONH₂, (CH₂)_(m)CONR₂, or (CH₂)_(m)CONHR; wherein R is H, alkylor acyl, and m is 0, 1 or 2;

wherein for base (B), W₄ cannot be CH if W₁, W₂, and W₃ are N; and

wherein for base (E), (F), (K), (L), (W), and (X), W₄ cannot be CH if W₁is N.

In some aspects of the present invention, compounds of Formula I arethose in which Z′ is —CHR²³—OH, C₁₋₄ cycloalkyl, C₂₋₄ alkenyl, or C₂₋₄alkynyl, wherein R²³ is methyl, ethyl, hydroxymethyl, fluoromethyl,—CH₂N₃, —CH₂—NR²¹R²², —CH₂—, or —CH₂—NH₂, wherein R²¹ and R²² areindependently H or lower alkyl. In other aspects, Z′, is —CHR²³—OH,—OC(R²³)₂PO₃H₂ or —CH₂C(R²³)₂PO₃H₂, wherein R²³ is methyl or ethyl.

In some aspects, Z′ is —O—, —CH(R²³)—O—, C₁₋₄ cycloalkylene, C₂₋₄alkenylene, or C₂₋₄ alkynylene; wherein R²³ is H, F, methyl, ethyl,hydroxymethyl, fluoromethyl, —CH₂N₃, —CH₂—NR²¹R²², —CH₂—, or —CH₂—NH₂,wherein R²¹ and R²² are independently H or lower alkyl.

In some aspects of the present invention, Z″ is absent. In otheraspects, Z″ is R²⁴(C═O)—, R²⁴—O—(C═O)—, or an ester of an L-amino acidsuch as an L-valine ester, e.g., R²⁴—CH(NH₂)(C═O)—, wherein R²⁴ isoptionally substituted C₁₋₆ alkyl, cycloalkyl, aryl, or aralkyl; or Z″is

wherein:

V, W, and W′ are independently H, optionally substituted alkyl,optionally substituted aralkyl, cycloalkyl, heterocycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, optionallysubstituted 1-alkenyl, or optionally substituted 1-alkynyl; and

Z is —CHR^(z)OH, —CHR^(z)OC(O)R^(y), —CH^(z)OC(S)R^(y),—CH^(z)OC(S)OR^(y), —CHR^(z)OC(O)SR^(y), —CHR^(z)OCO₂R^(y), —OR^(z),—SR^(z), —CHR^(z)N₃, —CH₂aryl, —CH(aryl)OH, —CH(CH═CR^(z) ₂)OH,—CH(C≡CR^(z))OH, —R^(z), —NR^(z) ₂, —OCOR^(y), —OCO₂R^(y), —SCOR^(y),—SCO₂R^(y), —NHCOR^(z), —NHCO₂R^(y), —CH₂NHaryl, —(CH₂)_(q)—OR^(z), and—(CH₂)_(q)—SR^(z), halogen, —CN, —COR^(y), —CONR^(z) ₂, —CO₂R^(y),—SO₂R^(y), or —SO₂NR^(z) ₂, wherein q is 2 or 3, R^(W) is R^(y) or —H,and R^(y) is alkyl, aryl, cycloalkyl, heterocycloalkyl, or aralkyl.

In some aspects of the present invention, Z″ is

wherein

V, Z, W, W′, q, R^(z), and R^(y) are as defined above.

In some aspects of the invention in which Z″ is

V and Z are connected together via an additional 3-5 atoms to form acyclic group containing 5-7 atoms, wherein 0-1 atoms are heteroatoms andthe remaining atoms are carbon substituted with hydroxy, acyloxy,alkylthiocarbonyloxy, alkoxycarbonyloxy, or aryloxycarbonyloxy attachedto a carbon atom that is three atoms from both O groups attached to thephosphorus.

In other aspects, V and Z are connected together via an additional 3-5atoms to form a cyclic group, wherein 0-1 atoms are heteroatoms and theremaining atoms are carbon, that is fused to an aryl group at the betaand gamma position to the O attached to the phosphorus. In yet otheraspects, V and W are connected together via an additional 3 carbon atomsto form an optionally substituted cyclic group containing 6 carbon atomsand substituted with one substituent selected from the group consistingof hydroxy, acyloxy, alkoxycarbonyloxy, alkylthiocarbonyloxy, andaryloxycarbonyloxy, attached to one of said carbon atoms that is threeatoms from an O attached to the phosphorus.

In other aspects, Z and W are connected together via an additional 3-5atoms to form a cyclic group, wherein 0-1 atoms are heteroatoms and theremaining atoms are carbon, and V must be aryl, substituted aryl,heteroaryl, or substituted heteroaryl.

Or, in yet other aspects, W and W′ are connected together via anadditional 2-5 atoms to form a cyclic group, wherein 0-2 atoms areheteroatoms and the remaining atoms are carbon, and V must be aryl,substituted aryl, heteroaryl, or substituted heteroaryl.

In those aspects of the invention wherein Z″ is

the following provisos apply:

a) V, Z, W, W′ are not all —H; and

b) when Z is —R^(z), then at least one of V, W, and W′ is not —H, alkyl,aralkyl, cycloalkyl or heterocycloalkyl.

In some aspects of the present invention, V is selected from the groupconsisting of phenyl; substituted phenyl with 1-3 substituentsindependently selected from the group consisting of halogen, C₁₋₆ alkyl,—CF₃, —OR³, —OR¹², —COR³, —CO₂R³, —N(R³)₂, —N(R¹²)₂, —CO₂N(R²)₂, —SR³,—SO₂R³, —SO₂N(R²)₂ and —CN; monocyclic heteroaryl; and substitutedmonocyclic heteroaryl with 1-2 substituents independently selected fromthe group consisting of halogen, C₁₋₆ alkyl, —CF₃, —OR₃, —OR¹², —COR³,—CO₂R³, —N(R³)₂, —N(R¹²)₂, —CO₂N(R²)₂, —SR³, —SO₂R³, —SO₂N(R¹²)₂ and—CN; wherein said monocyclic heteroaryl and substituted monocyclicheteroaryl has 1-2 heteroatoms that are independently selected from thegroup consisting of N, O, and S; wherein R² is H or R³, R³ is C₁₋₆alkyl, aryl, heterocycloalkyl, or aralkyl, and R¹² is H or lower acyl;with the provisos that

a) when there are two heteroatoms and one is 0, then the other can notbe O or S, and

b) when there are two heteroatoms and one is S, then the other can notbe or S; or

V and Z together are connected via an additional 3-5 atoms to form acyclic group, optionally containing 1 heteroatom, that is fused to anaryl group at the beta and gamma position to the O attached to thephosphorus.

In other aspects, V is selected from the group consisting of phenyl;substituted phenyl with 1-3 substituents independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, —CF₃, —COCH₃, —OMe, —NMe₂,—OEt, —CO₂t-butyl, —CO₂NH₂, —SMe, —SO₂Me, —SO₂NH₂, and —CN; monocyclicheteroaryl; and substituted monocyclic heteroaryl with 1-2 substituentsindependently selected from the group consisting of —Cl, —Br, —F, C₁₋₃alkyl, —CF₃, —COCH₃, —OMe, —NMe₂, —OEt, —CO₂t-butyl, —CO₂NH₂, —SMe,—SO₂Me, —SO₂NH₂ and —CN; wherein said monocyclic heteroaryl andsubstituted monocyclic heteroaryl has 1-2 heteroatoms that areindependently selected from the group consisting of N, O, and S, withthe provisos that

a) when there are two heteroatoms and one is 0, then the other can notbe O or S, and

b) when there are two heteroatoms and one is S, then the other can notbe O or S; or

V and Z are connected together via an additional 4 atoms to form a6-membered ring that is fused to a phenyl or substituted phenyl at thebeta and gamma position to the O attached to the phosphorus.

In yet other aspects of the present invention, V is selected from thegroup consisting of phenyl; substituted phenyl with 1-2 substituentsindependently selected from the group consisting of —Cl, —Br, —F, C₁₋₃alkyl, and —CF₃; pyridyl; substituted pyridyl with 1 substituentindependently selected from the group consisting of —Cl, —Br, —F, C₁₋₃alkyl, and —CF₃; furanyl; substituted furanyl with 1 substituentindependently selected from the group consisting of —Cl, —Br, —F, C₁₋₃alkyl, and —CF₃; thienyl; and substituted thienyl with 1 substituentindependently selected from the group consisting of —Cl, —Br, —F, C₁₋₃alkyl, and —CF₃.

In further aspects, V is selected from the group consisting of phenyl,3-chlorophenyl, 3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl,3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl, and 4-pyridyl. In otheraspects, V is selected from the group consisting of 3-chlorophenyl,3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl, 3-pyridyl, and4-pyridyl.

In some aspects, V is selected from the group consisting of phenyl;substituted phenyl with 1-3 substituents independently selected from thegroup consisting of —Cl, —Br, —P, C₁₋₃ alkyl, —CF₃, —COCH₃, —OH, —OMe,—NH₂, —NMe₂, —OEt, —COOH, —CO₂t-butyl, —CO₂NH₂, —SMe, —SO₂Me, —SO₂NH₂and —CN; monocyclic heteroaryl; and substituted monocyclic heteroarylwith 1-2 substituents independently selected from the group consistingof —Cl, —Br, —F, C₁₋₃ alkyl, —CF₃, —COCH₃, —OH, —OMe, —NH₂, —NMe₂, —OEt,—COOH, —CO₂t-butyl, —CO₂NH₂, —SMe, —SO₂Me, —SO₂NH₂ and —CN; wherein saidmonocyclic heteroaryl and substituted monocyclic heteroaryl has 1-2heteroatoms that are independently selected from the group consisting ofN, O, and S; with the provisos that

a) when there are two heteroatoms and one is O, then the other can notbe O or S, and

b) when there are two heteroatoms and one is S, then the other can notbe O or S; or

V and Z are connected together via an additional 4 atoms to form a6-membered ring that is fused to a phenyl or substituted phenyl at thebeta and gamma position to the 0 attached to the phosphorus.

In some aspects, Z is selected from the group consisting of —H, —OMe,—OEt, phenyl, C₁₋₃ alkyl, —N(R⁴)₂, —SR⁴, —(CH₂)_(p)—OR⁶, —(CH₂)_(p)—SR⁶and —OCOR⁵; wherein R⁴ is C₁-C₄ alkyl; R⁵ is selected from the groupconsisting of C₁-C₄ alkyl, monocyclic aryl, and monocyclic aralkyl; andR⁶ is C₁-C₄ acyl. In further aspects, Z is selected from the groupconsisting of H, —OMe, —OEt, and phenyl.

In some aspects, W and W′ are independently selected from the groupconsisting of H, C₁₋₆ alkyl, and phenyl; or W and W′ are connectedtogether via an additional 2-5 atoms to form a cyclic group. In yetother aspects, W and W′ are independently selected from the groupconsisting of H, methyl, and V, or W and W′ are each methyl, with theproviso that when W is V, then W′ is H.

In some aspects, V is selected from the group consisting of optionallysubstituted monocyclic aryl and optionally substituted monocyclicheteroaryl;

W and W′ are independently selected from the group consisting of —H,methyl, and V; or W and W′ are each methyl; with the proviso that when Wis V, then W′ is H; and

Z is selected from the group consisting of —H, —OMe, —OEt, phenyl, C₁₋₃alkyl, —N(R⁴)₂, —SR⁴, —(CH₂)_(p)—OR⁶, —(CH₂)_(p)—SR⁶ and —OCOR⁵, whereinR⁴ is C₁₋₄ alkyl; R⁵ is selected from the group consisting of C₁₋₄alkyl, monocyclic aryl, and monocyclic aralkyl; and R⁶ is C₁₋₄ acyl; or

Z and V are connected together via an additional 3-5 atoms to form acyclic group, optionally containing 1 heteroatom, that is fused to anaryl group at the beta and gamma position to the O attached to thephosphorus; or

Z and W are connected together via an additional 3-5 atoms to form acyclic group, optionally containing one heteroatom; or

W and W′ are connected together via an additional 2-5 atoms to form acyclic group.

In other aspects, V is selected from the group consisting of phenyl;substituted phenyl with 1-3 substituents independently selected from thegroup consisting of halogen, C₁₋₆ alkyl, —CF₃, —OR³, —OR¹², —COR³,—CO₂R³, —N(R³)₂, —N(R¹²)₂, —CO₂N(R²)₂, —SR³, —SO₂R³, —SO₂N(R²)₂ and —CN;monocyclic heteroaryl; and substituted monocyclic heteroaryl with 1-2substituents independently selected from the group consisting ofhalogen, C₁₋₆ alkyl, —CF₃, OR³, —OR¹², —COR³, —CO₂R³, —N(R¹²)₂,—CO₂N(R²)₂, —SR³, —SO₂R³, —SO₂N(R²)₂ and —CN; wherein said monocyclicheteroaryl and substituted monocyclic heteroaryl has 1-2 heteroatomsthat are independently selected from the group consisting of N, O, andS; wherein R² is H or R³, R³ is C₁₋₆ alkyl, aryl, heterocycloalkyl, oraralkyl, and R¹² is H or lower acyl; with the provisos that

a) when there are two heteroatoms and one is O, then the other can notbe O or S, and

b) when there are two heteroatoms and one is S, then the other can notbe O or S; or

W and W′ are independently selected from the group consisting of —H,methyl, and V; or W and W′ are each methyl, with the proviso that when Wis V, then W′ is H;

Z is selected from the group consisting of —H, —OMe, —OEt, phenyl, C₁-C₃alkyl, —N(R⁴)₂, —SR⁴, —(CH₂)_(p)—OR⁶, —(CH₂)_(p)—SR⁶ and —OCOR⁵; whereinR⁴ is C₁-C₄ alkyl, R⁵ is selected from the group consisting of C₁-C₄alkyl, monocyclic aryl, and monocyclic aralkyl, and R⁶ is C₁-C₄ acyl; or

Z and V are connected together via an additional 3-5 atoms to form acyclic group, optionally containing 1 heteroatom, that is fused to anaryl group at the beta and gamma position to the 0 attached to thephosphorus; or

Z and W are connected together via an additional 3-5 atoms to form acyclic group, optionally containing one heteroatom; or

W and W′ are connected together via an additional 2-5 atoms to form acyclic group.

In other aspects, V is selected from the group consisting of phenyl;substituted phenyl with 1-3 substituents independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, —CF₃, —COCH₃, —OMe, —NMe₂,—OEt, —CO₂t-butyl, —CO₂NH₂, —SMe, —SO₂Me, —SO₂NH₂ and —CN; monocyclicheteroaryl; and substituted monocyclic heteroaryl with 1-2 substituentsindependently selected from the group consisting of —Cl, —Br, —F, C₁₋₃alkyl, —CF₃, —COCH₃, —OMe, —NMe₂, —OEt, —CO₂t-butyl, —CO₂NH₂, —SMe,—SO₂Me, —SO₂NH₂ and —CN; wherein said monocyclic heteroaryl andsubstituted monocyclic heteroaryl has 1-2 heteroatoms that areindependently selected from the group consisting of N, O, and S; withthe provisos that

a) when there are two heteroatoms and one is O, then the other can notbe O or S; and

b) when there are two heteroatoms and one is S, then the other can notbe O or S; or

W and W′ are independently selected from the group consisting of —H,methyl, and V, or W and W′ are each methyl, with the proviso that when Wis V, then W′ is H;

Z is selected from the group consisting of —H, —OMe, —OEt, phenyl, C₁₋₃alkyl, —N(R⁴)₂, —SR⁴, —(CH₂)_(p)—OR⁶, —(CH₂)_(p)—SR⁶ and —OCOR⁵, whereinR⁴ is C₁-C₄ alkyl, R⁵ is selected from the group consisting of C₁-C₄alkyl, monocyclic aryl, and monocyclic aralkyl, and R⁶ is C₁-C₄ acyl; or

V and Z are connected together via an additional 4 atoms to form a6-membered ring that is fused to a phenyl or substituted phenyl at thebeta and gamma position to the 0 attached to the phosphorus; or

Z and W are connected together via an additional 3-5 atoms to form acyclic group, optionally containing one heteroatom; or

W and W′ are connected together via an additional 2-5 atoms to form acyclic group.

In yet other aspects, V is selected from the group consisting of phenyl;substituted phenyl with 1-2 substituents independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, and —CF₃; pyridyl;substituted pyridyl with 1 substituent independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, and —CF₃; furanyl;substituted furanyl with 1 substituent independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, and —CF₃; thienyl; andsubstituted thienyl with 1 substituent independently selected from thegroup consisting of —Cl, —Br, —F, C₁-C₃ alkyl, and —CF₃;

W and W′ are independently selected from the group consisting of —H,methyl, and V, or W and W′ are each methyl, with the proviso that when Wis V, then W′ is H; and

Z is selected from the group consisting of —H, —OMe, —OEt, phenyl, C₁-C₃alkyl, —N(R⁴)₂, —SR⁴, —(CH₂)_(p)—OR⁶, —(CH₂)_(p)—SR⁶ and —OCOR⁵, whereinR⁴ is C₁-C₄ alkyl, R⁵ is selected from the group consisting of C₁-C₄alkyl, monocyclic aryl, and monocyclic aralkyl, and R⁶ is C₁-C₄ acyl; or

Z and W are connected together via an additional 3-5 atoms to form acyclic group, optionally containing one heteroatom; or

W and W′ are connected together via an additional 2-5 atoms to form acyclic group.

In further aspects, V is selected from the group consisting of phenyl,3-chlorophenyl, 3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl,3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl, and 4-pyridyl;

Z is selected from the group consisting of —H, OMe, OEt, and phenyl; and

W and W′ are independently selected from the group consisting of —H andphenyl, or W and W′ are each methyl.

In some aspects, Z, W, and W′ are each —H. In other aspects, V and W arethe same and each is selected from the group consisting of optionallysubstituted monocyclic aryl and optionally substituted monocyclicheteroaryl.

In some aspects of the present invention, Z″ is:

wherein

V is as defined above.

In other aspects, Z″ is selected from the following non-limitingexamples:

In some aspects of the present invention, Z″ is P(O)Y′R¹¹Y″R¹¹, whereineach R¹¹ is independently H or C₁₋₄ alkyl;

Y′ and Y″ are each independently selected from the group consisting of—O—, and —NR^(v)—; and

when Y′ and Y″ are both —O—, R¹¹ attached to —O— is independentlyselected from the group consisting of optionally substituted aryl,optionally substituted CH₂-heterocycloakyl wherein the cyclic moietycontains a carbonate or thiocarbonate, optionally substituted-alkylaryl, —C(z)₂OC(O)NR^(z) ₂, —NR^(z)—C(O)—R^(y),—C(R^(z))₂—OC(O)R^(Y), —C(R^(z))₂—O—C(O)OR^(y), —C(R^(z))₂OC(O)SR^(y),-alkyl-S—C(O)R^(y), -alkyl-S—S-alkylhydroxy, and-alkyl-S—S—S-alkylhydroxy; or

when Y′ and Y″ are both —NR^(v)—, then R¹¹ attached to —NR^(v)— isindependently selected from the group consisting of —H,—[C(R^(z))₂]_(q)—COOR^(y), —C(R^(x))₂COOR^(y),-[C(R^(z))₂]_(q)—C(O)SR^(y), and -cycloalkylene-COOR^(y); or

when Y′ is —O— and Y″ is NR^(v), then R¹¹ attached to —O— isindependently selected from the group consisting of optionallysubstituted aryl, optionally substituted CH₂-heterocycloakyl wherein thecyclic moiety contains a carbonate or thiocarbonate, optionallysubstituted -alkylaryl, —C(R^(z))₂OC(O)NR^(z) ₂, —NR^(z)—C(O)—R^(y),—C(R^(z))₂—OC(O)R^(y), —C(R^(z))₂—O—C(O)OR^(y), —C(R^(z))₂OC(O)SR^(y),-alkyl-S—C(O)R^(y), -alkyl-S—S-alkylhydroxy, and-alkyl-S—S—S-alkylhydroxy; and R¹¹ attached to —NR^(v)— is independentlyselected from the group consisting of —H, —[C(R^(z))₂]_(q)—COOR^(y),—C(R^(x))₂COOR^(y), —[C(R^(z))₂]_(q)—C(O)SR^(y), and-cycloalkylene-COOR^(y); or

when Y′ and Y″ are independently selected from —O— and —NR^(v)—, thenR¹¹ and R¹¹ together form a cyclic group comprising -alkyl-S—S-alkyl-;

wherein q is an integer 2 or 3;

each R^(z) is selected from the group consisting of R^(y) and —H;

each R^(y) is selected from the group consisting of alkyl, aryl,heterocycloalkyl, and aralkyl;

each R^(x) is independently selected from the group consisting of —H,and alkyl, or together R^(x) and R^(x) form a cycloalkyl group; and

each R^(v) is selected from the group consisting of —H, lower alkyl,acyloxyalkyl, alkoxycarbonyloxyalkyl, and lower acyl.

In one aspect, Z″ is —P(O)Y′R¹¹Y″R¹¹.

In one aspect, Z″ is selected from the group consisting of—P(O)[—OCR^(z) ₂OC(O)R^(y)]₂, —P(O)[—OCR^(z) ₂OC(O)OR^(y)]₂,—P(O)[—N(H)CR^(z) ₂C(O)OR^(y)]₂, —P(O)[—N(H)CR^(z) ₂C(O)OR^(y)][—OR¹¹],

In another aspect, Z″ is selected from the group consisting of—P(O)[—OCR^(z) ₂OC(O)R^(y)]₂, —P(O)[—OCR^(z) ₂OC(O)OR^(y)]₂,—P(O)[—OCH₂CH₂SC(O)Me]₂, —P(O)[—N(H)CR^(z) ₂C(O)OR^(y)]₂, and—P(O)[—N(H)CR^(z) ₂C(O)OR^(y)][—OR¹¹].

In another aspect, Z″ is selected from the group consisting of—P(O)[—OCR^(z) ₂OC(O)R^(y)]₂, —P(O)[—OCR^(z) ₂OC(O)OR^(y)]₂,—P(O)[—Oalk-SC(O)R^(y)]₂, —P(O)[—N(H)CR^(z) ₂C(O)OR^(y)]₂, and—P(O)[—N(H)CR^(z) ₂C(O)OR^(y)][—OR¹¹].

In one aspect, Z″ is selected from the group consisting of—P(O)[—OCR^(z) ₂OC(O)R^(y)]₂, —P(O)[—OCR^(z) ₂OC(O)OR^(y)]₂,—P(O)[—N(H)CR^(z) ₂C(O)OR^(y)]₂, —P(O)[—N(H)CR^(z) ₂C(O)OR^(y)][—OR¹¹],—P(O)(OH)(OR¹¹), —P(O)(OR^(e))(OR^(e), —P(O)[—OCR^(z)₂OC(O)R^(y)](OR^(e)), —P(O)[—OCR^(z) ₂OC(O) OR^(y)](OR^(e)), and—P(O)[—N(H)CR^(z) ₂C(O)OR^(y)](OR^(e)),

In another aspect, Z″ is selected from the group consisting of—P(O)[—OCR_(z) ²OC(O)R^(y)]₂, —P(O)[—OCR^(z) ₂OC(O)OR^(y)]₂,—P(O)[—N(H)CR^(z) ₂C(O)OR^(y)]₂, —P(O)[—N(H)CR^(z) ₂C(O)OR^(y)][—OR¹¹],—P(O)(OH)(OR^(e)), —P(O)(OR^(e))(OR^(e)), —P(O)[—OCR^(z)₂OC(O)R^(y)](OR^(e)), —P(O)[—O CR^(z) ₂OC(O)OR^(y)](OR^(e)),—P(O)[—N(H)CR^(z) ₂C(O)OR^(y)](OR^(e)), and

In one aspect, Z″ is selected from the group consisting of—P(O)[—OCH₂OC(O)-t-butyl]₂, —P(O)[—OCH₂OC(O)O-1-propyl]₂,—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃]₂, —P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃]₂,—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃][3,4-methylenedioxyphenyl],—P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃][3,4-methylenedioxyphenyl],—P(O)[—O—CH₂CH₂S—C(O)CH₃]₂, and —P(O)[—OCH(3-chlorophenyl)CH₂CH₂O—]. Ina further aspect, Z″ is selected from the group consisting of—P(O)[—OCH₂OC(O)-t-butyl]₂, —P(O)[—OCH₂OC(O)O-i-propyl]₂,—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃]₂, —P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃]₂,—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃][3,4-methylenedioxy-phenyl],—P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃][3,4-methylenedioxyphenyl], and—P(O)[—OCH(3-chlorophenyl)CH₂CH₂O—].

In yet another aspect, Z″ is selected from the group consisting of—P(O)[—OCH₂OC(O)-t-butyl]₂ and —P(O)[—OCH₂OC(±)-1-propyl]₂. In anotheraspect, Z″ is selected from the group consisting of—P(O)[—OCH₂OC(O)-t-butyl]₂, —P(O)[—OCH₂OC(O)O-1-propyl]₂,—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃]₂, —P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃]₂,—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃][3,4-methylenedioxyphenyl],—P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃][3,4-methylenedioxyphenyl],—P(O)[—OCH₂OC(O)-t-butyl] (OCH₃), —P(O)[—OCH₂OC(O)O-1-propyl](OCH₃),—P(O)[—OCH(CH₃)OC(O)-t-butyl] (OCH₃),—P(O)[—OCH(CH₃)OC(O)O-i-propyl](OCH₃),—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃](OCH₃),—P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃](OCH₃), and —P(O)(OH)(NH₂).

In one aspect, Z″ is selected from the group consisting of—P(O)[—OCH₂OC(O)O-ethyl]₂ and —P(O)[—OCH₂OC(O)O-1-propyl]₂. In anotheraspect, Z″ is selected from the group consisting of—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃]₂ and —P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃]₂—In afurther aspect, Z″ is —P(O)[—OCH₂CH₂SC(O)Me]₂. In another aspect, Z″ isselected from the group consisting of—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃][3,4-methylenedioxyphenyl] and—P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃][3,4-methylenedioxyphenyl]. In a furtheraspect, Z″ is selected from the group consisting of —P(O)[—OCR^(z)₂OC(O)R^(y)]₂, —P(O)[—OCR^(z) ₂OC(O)OR^(y)]₂, —P(O)[—N(H)CR^(z)₂C(O)OR^(y)]₂, —P(O)[—N(H)CR^(z) ₂C(O)OR^(y)][—OR¹¹ ] and —P(O)[—OCH(V)CH₂CH₂O—]. In another aspect, Z″ is selected from the group consistingof —P(O)[—OCH₂OC(O)-t-butyl]₂, —P(O)[—OCH₂OC(O)O-i-propyl]₂,—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃]₂, —P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃]₂,—P(O)[—N(H)CH(CH₃)C(O)OCH₂CH₃][3,4-methylenedioxyphenyl],—P(O)[—N(H)C(CH₃)₂C(O)OCH₂CH₃] [3,4-methylenedioxyphenyl], and—P(O)[—OCH(3-chlorophenyl)CH₂CH₂O—].

In some aspects, the present invention relates to compounds of FormulaeII-VIII, and hydrates, solvates, prodrugs, co-crystals, andpharmaceutically acceptable salts thereof, including stereoisomersthereof and mixtures of stereoisomers thereof:

wherein X′, Y, B, R¹⁵, R¹⁶, R¹⁷, R¹⁸, R¹⁹, Z′, Z″, V, Z, W, and W′ areas defined above for Formula I.

Some of the compounds of Formulae I-VIII have asymmetric centers wherethe stereochemistry is unspecified, and the diastereomeric mixtures ofthese compounds are included, as well as the individual stereoisomerswhen referring to a compound of Formulae I-VIII generally.

In some aspects, Formulae II-VIII have the stereochemistry of Formula I.

For example, compounds of Formula II above include compounds with thefollowing structure:

Some of the compounds described herein may also exist as tautomers suchas keto-enol tautomers and imine-enamine tautomers. The individualtautomers as well as mixtures thereof are encompassed with compounds ofFormulae I-VIII. An example of keto-enol tautomers which are intended tobe encompassed within the compounds of the present invention isillustrated below:

An example of imine-enamine tautomers which are intended to beencompassed within the compounds of the present invention is illustratedbelow:

A further aspect of this invention includes compounds of Formula XVI andisomers, solvates, hydrates, prodrugs, or pharmaceutically acceptablesalts thereof:

wherein:

B and X′ are as defined in Formula I above;

V is selected from the group consisting of optionally substitutedmonocyclic aryl and optionally substituted monocyclic heteroaryl; and

V and the 5′ oxymethylene group of the ribose sugar moiety are cis toone another.

In further aspects, this invention includes a compound of Formula XVI,or a solvate, hydrate, prodrug, or pharmaceutically acceptable saltthereof, wherein:

B and X′ are as defined in Formula I above;

V is selected from the group consisting of optionally substitutedmonocyclic aryl and optionally substituted monocyclic heteroaryl; and

V and the 5′ oxymethylene group of the ribose sugar moiety are cis toone another.

In additional aspects, compounds of Formula XVI are those in which V isselected from the group consisting of phenyl; substituted phenyl with1-3 substituents independently selected from the group consisting ofhalogen, C₁₋₆ alkyl, —CF₃, —OR³, —OR¹², —COR³, —CO₂R³, —N(R³)₂,—N(R¹²)₂, —CO₂N(R²)₂, —SR³, —SO₂R³, —SO₂N(R²)₂ and —CN; monocyclicheteroaryl; and substituted monocyclic heteroaryl with 1-2 substituentsindependently selected from the group consisting of halogen, C₁₋₆ alkyl,—CF₃, —OR³, —OR¹², —COR³, —CO₂R³, —N(R³)₂, —N(R¹²)₂, —CO₂N(R²)₂, —SR³,—SO₂R³, —SO₂N(R²)₂ and —CN; wherein R³ is C₁-C₆ alkyl, and R¹² is H andC₁-C₆ alkyl, and wherein said monocyclic heteroaryl and substitutedmonocyclic heteroaryl has 1-2 heteroatoms that are independentlyselected from the group consisting of N, O, and S; with the provisosthat

a) when there are two heteroatoms and one is O, then the other can notbe O or S, and

b) when there are two heteroatoms and one is S, then the other can notbe O or S.

In further aspects, V of Formula XVI is selected from the groupconsisting of phenyl; substituted phenyl with 1-3 substituentsindependently selected from the group consisting of —Cl, —Br, —F, C₁₋₃alkyl, —CF₃, —COCH₃, —OMe, —NMe₂, —OEt, —CO₂t-butyl, —CO₂NH₂, —SMe,—SO₂Me, —SO₂NH₂ and —CN; monocyclic heteroaryl; and substitutedmonocyclic heteroaryl with 1-2 substituents independently selected fromthe group consisting of —Cl, —Br, —F, C₁₋₃ alkyl, —CF₃, —COCH₃, —OMe,—NMe₂, —OEt, —CO₂t-butyl, —CO₂NH₂, —SMe, —SO₂Me, —SO₂NH₂ and —CN;wherein said monocyclic heteroaryl and substituted monocyclic heteroarylhas 1-2 heteroatoms that are independently selected from the groupconsisting of N, O, and S; with the provisos that

a) when there are two heteroatoms and one is O, then the other can notbe O or S, and

b) when there are two heteroatoms and one is S, then the other can notbe O or S; or

V and Z are connected together via an additional 4 atoms to form a6-membered ring that is fused to a phenyl or substituted phenyl at thebeta and gamma position to the O attached to the phosphorus.

In additional aspects, V of Formula XVI is selected from the groupconsisting of phenyl; substituted phenyl with 1-2 substituentsindependently selected from the group consisting of —Cl, —Br, —F, C₁-C₃alkyl, and —CF₃; pyridyl; substituted pyridyl with 1 substituentindependently selected from the group consisting of —Cl, —Br, —F, C₁-C₃alkyl, and —CF₃; furanyl; substituted furanyl with 1 substituentindependently selected from the group consisting of —Cl, —Br, —F, C₁-C₃alkyl, and —CF₃; thienyl; and substituted thienyl with 1 substituentindependently selected from the group consisting of —Cl, —Br, —F, C₁-C₃alkyl, and —CF₃.

In yet other aspects, V is selected from the group consisting of phenyl,3-chlorophenyl, 3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl,3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl, and 4-pyridyl. In otheraspects, V is selected from the group consisting of 3-chlorophenyl,3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl, 3-pyridyl, and4-pyridyl.

In some aspects, V of Formula XVI is selected from the group consistingof phenyl; substituted phenyl with 1-3 substituents independentlyselected from the group consisting of —Cl, —Br, —F, C₁₋₃ alkyl, —CF₃,—COCH₃, —OMe, —NMe₂, —OEt, —CO₂t-butyl, —CO₂NH₂, —SMe, —SO₂Me, —SO₂NH₂and —CN; monocyclic heteroaryl; and substituted monocyclic heteroarylwith 1-2 substituents independently selected from the group consistingof —Cl, —Br, —F, C₁-C₃ alkyl, —CF₃, —COCH₃, —OMe, —NMe₂, —OEt,—CO₂t-butyl, —CO₂NH₂, —SMe, —SO₂Me, —SO₂NH₂ and —CN. In other aspects, Vis selected from the group consisting of phenyl, 3-chlorophenyl,3-bromophenyl, 2-bromophenyl, 3,5-dichlorophenyl,3-bromo-4-fluorophenyl, 2-pyridyl, 3-pyridyl, and 4-pyridyl.

In some aspects of the present invention, the compounds of thisinvention have R-stereochemistry at the V-attached carbon and haveS-stereochemistry at the phosphorus center. In other aspects, thecompounds of this invention have S-stereochemistry at the V-attachedcarbon and have R-stereochemistry at the phosphorus center.

In some aspects, the following compounds are included in the inventionbut the compounds are not limited to these illustrative compounds.

The following prodrugs are preferred compounds of the invention.

The compounds are shown without depiction of stereochemistry since thecompounds are biologically active as the diastereomeric mixture or as asingle stereoisomer. Compounds named in Table 1 are designated bynumbers assigned to the variables of formula using the followingconvention: M1.V.L1.L2. M1 is a variable that represents nucleosides ofFormulae IX-XIV which are attached via 5′-hydroxyl group that isphosphorylated with a group P(O)(O—CH(V)CH₂CH₂—O) to make compounds ofFormula XV. V is an aryl or heteroaryl group that has 2 substituents, L1and L2, at the designated positions. V may have additional substituents.

Variable M1:

wherein Y, R¹⁹, R¹⁸, R¹⁷, R¹⁶, R¹⁵, X′ and B are as defined above forFormula I.

Variable V: Group V1

1) 2-(L1)-3(L2)-phenyl

2) 2-(L1)-4(L2)-phenyl

3) 2-(L1)-5(L2)-phenyl

4) 2-(L1)-6(L2)-phenyl

5) 3-(L1)-4(L2)-phenyl

6) 3-(L1)-5(L2)-phenyl

7) 3-(L1)-6(L2)-phenyl

8) 2-(L1)-6(L2)-3-chlorophenyl

9) 4-(L1)-5(L2)-3-chlorophenyl

Variable V: Group V2

1) 2-(L1)-3(L2)-4-pyridyl

2) 2-(L1)-5(L2)-4-pyridyl

3) 2-(L1)-6(L2)-4-pyridyl

4) 3-(L1)-5(L2)-4-pyridyl

5) 3-(L1)-6(L2)-4-pyridyl

6) 2-(L1)-4(L2)-3-pyridyl

7) 2-(L1)-5(L2)-3-pyridyl

8) 2-(L1)-6(L2)-3-pyridyl

9) 4-(L1)-5(L2)-3-pyridyl

Variable V: Group V3

1) 4-(L1)-6(L2)-3-pyridyl

2) 5-(L1)-6(L2)-3-pyridyl

3) 3-(L1)-4(L2)-2-pyridyl

4) 3-(L1)-5(L2)-2-pyridyl

5) 3-(L1)-6(L2)-2-pyridyl

6) 4-(L1)-5(L2)-2-pyridyl

7) 4-(L1)-6(L2)-2-pyridyl

8) 3-(L1)-4(L2)-2-thienyl

9) 3-(L1)-4(L2)-2-furanyl

Variable L1

1) hydrogen

2) chloro

3) bromo

4) fluoro

5) methyl

6) trifluoromethyl

7) methoxy

8) dimethylamino

9) cyano

Variable L2

1) hydrogen

2) chloro

3) bromo

4) fluoro

5) methyl

6) trifluoromethyl

7) methoxy

8) dimethylamino

9) cyano

Preferred groups of compounds are those listed in Table 1 usingvariables M1 and V1 and L1 and L2 listed in that order. For example,“1.3.6.7” represents structure 1 of variable Ml (for example, where Y isO, R¹⁹ is absent, R¹⁵—R¹⁸ are each H, and B is 7-deaza-2′-methyladenine, the 2′,3′-cyclic carbonate form of 7-deaza-2′-methyladenosine); structure 3 of group V1 (i.e., 2-(L1)-5-(L2) phenyl);structure 6 of variable L1 (i.e., trifluoromethyl); and structure 7 ofvariable L2 (i.e., methoxy). The group 1.3.6.7. therefore includes7-deaza-2′-methyladenosine 2′,3′-cyclic carbonate with theP(O)(O—CH(V)CH₂CH₂O) group attached to the 5′-primary hydroxyl of theribose ring being[1-(2-trifluoromethyl-5-methoxyphenyl)-1,3-propyl]phosphoryl.

Preferred groups of compounds are also [those listed in Table 1 usingvariables M1 and V2 wherein the four digit number representsM1.V2.L1.L2.

Preferred groups of compounds are also those listed in Table 1 usingvariables M1 and V3 wherein the four digit number representsM1.V3.L1.L2.

TABLE 1 1.1.1.1 1.1.1.2 1.1.1.3 1.1.1.4 1.1.1.5 1.1.1.6 1.1.1.7 1.1.1.81.1.1.9 1.1.2.1 1.1.2.2 1.1.2.3 1.1.2.4 1.1.2.5 1.1.2.6 1.1.2.7 1.1.2.81.1.2.9 1.1.3.1 1.1.3.2 1.1.3.3 1.1.3.4 1.1.3.5 1.1.3.6 1.1.3.7 1.1.3.81.1.3.9 1.1.4.1 1.1.4.2 1.1.4.3 1.1.4.4 1.1.4.5 1.1.4.6 1.1.4.7 1.1.4.81.1.4.9 1.1.5.1 1.1.5.2 1.1.5.3 1.1.5.4 1.1.5.5 1.1.5.6 1.1.5.7 1.1.5.81.1.5.9 1.1.6.1 1.1.6.2 1.1.6.3 1.1.6.4 1.1.6.5 1.1.6.6 1.1.6.7 1.1.6.81.1.6.9 1.1.7.1 1.1.7.2 1.1.7.3 1.1.7.4 1.1.7.5 1.1.7.6 1.1.7.7 1.1.7.81.1.7.9 1.1.8.1 1.1.8.2 1.1.8.3 1.1.8.4 1.1.8.5 1.1.8.6 1.1.8.7 1.1.8.81.1.8.9 1.1.9.1 1.1.9.2 1.1.9.3 1.1.9.4 1.1.9.5 1.1.9.6 1.1.9.7 1.1.9.81.1.9.9 1.2.1.1 1.2.1.2 1.2.1.3 1.2.1.4 1.2.1.5 1.2.1.6 1.2.1.7 1.2.1.81.2.1.9 1.2.2.1 1.2.2.2 1.2.2.3 1.2.2.4 1.2.2.5 1.2.2.6 1.2.2.7 1.2.2.81.2.2.9 1.2.3.1 1.2.3.2 1.2.3.3 1.2.3.4 1.2.3.5 1.2.3.6 1.2.3.7 1.2.3.81.2.3.9 1.2.4.1 1.2.4.2 1.2.4.3 1.2.4.4 1.2.4.5 1.2.4.6 1.2.4.7 1.2.4.81.2.4.9 1.2.5.1 1.2.5.2 1.2.5.3 1.2.5.4 1.2.5.5 1.2.5.6 1.2.5.7 1.2.5.81.2.5.9 1.2.6.1 1.2.6.2 1.2.6.3 1.2.6.4 1.2.6.5 1.2.6.6 1.2.6.7 1.2.6.81.2.6.9 1.2.7.1 1.2.7.2 1.2.7.3 1.2.7.4 1.2.7.5 1.2.7.6 1.2.7.7 1.2.7.81.2.7.9 1.2.8.1 1.2.8.2 1.2.8.3 1.2.8.4 1.2.8.5 1.2.8.6 1.2.8.7 1.2.8.81.2.8.9 1.2.9.1 1.2.9.2 1.2.9.3 1.2.9.4 1.2.9.5 1.2.9.6 1.2.9.7 1.2.9.81.2.9.9 1.3.1.1 1.3.1.2 1.3.1.3 1.3.1.4 1.3.1.5 1.3.1.6 1.3.1.7 1.3.1.81.3.1.9 1.3.2.1 1.3.2.2 1.3.2.3 1.3.2.4 1.3.2.5 1.3.2.6 1.3.2.7 1.3.2.81.3.2.9 1.3.3.1 1.3.3.2 1.3.3.3 1.3.3.4 1.3.3.5 1.3.3.6 1.3.3.7 1.3.3.81.3.3.9 1.3.4.1 1.3.4.2 1.3.4.3 1.3.4.4 1.3.4.5 1.3.4.6 1.3.4.7 1.3.4.81.3.4.9 1.3.5.1 1.3.5.2 1.3.5.3 1.3.5.4 1.3.5.5 1.3.5.6 1.3.5.7 1.3.5.81.3.5.9 1.3.6.1 1.3.6.2 1.3.6.3 1.3.6.4 1.3.6.5 1.3.6.6 1.3.6.7 1.3.6.81.3.6.9 1.3.7.1 1.3.7.2 1.3.7.3 1.3.7.4 1.3.7.5 1.3.7.6 1.3.7.7 1.3.7.81.3.7.9 1.3.8.1 1.3.8.2 1.3.8.3 1.3.8.4 1.3.8.5 1.3.8.6 1.3.8.7 1.3.8.81.3.8.9 1.3.9.1 1.3.9.2 1.3.9.3 1.3.9.4 1.3.9.5 1.3.9.6 1.3.9.7 1.3.9.81.3.9.9 1.4.1.1 1.4.1.2 1.4.1.3 1.4.1.4 1.4.1.5 1.4.1.6 1.4.1.7 1.4.1.81.4.1.9 1.4.2.1 1.4.2.2 1.4.2.3 1.4.2.4 1.4.2.5 1.4.2.6 1.4.2.7 1.4.2.81.4.2.9 1.4.3.1 1.4.3.2 1.4.3.3 1.4.3.4 1.4.3.5 1.4.3.6 1.4.3.7 1.4.3.81.4.3.9 1.4.4.1 1.4.4.2 1.4.4.3 1.4.4.4 1.4.4.5 1.4.4.6 1.4.4.7 1.4.4.81.4.4.9 1.4.5.1 1.4.5.2 1.4.5.3 1.4.5.4 1.4.5.5 1.4.5.6 1.4.5.7 1.4.5.81.4.5.9 1.4.6.1 1.4.6.2 1.4.6.3 1.4.6.4 1.4.6.5 1.4.6.6 1.4.6.7 1.4.6.81.4.6.9 1.4.7.1 1.4.7.2 1.4.7.3 1.4.7.4 1.4.7.5 1.4.7.6 1.4.7.7 1.4.7.81.4.7.9 1.4.8.1 1.4.8.2 1.4.8.3 1.4.8.4 1.4.8.5 1.4.8.6 1.4.8.7 1.4.8.81.4.8.9 1.4.9.1 1.4.9.2 1.4.9.3 1.4.9.4 1.4.9.5 1.4.9.6 1.4.9.7 1.4.9.81.4.9.9 1.5.1.1 1.5.1.2 1.5.1.3 1.5.1.4 1.5.1.5 1.5.1.6 1.5.1.7 1.5.1.81.5.1.9 1.5.2.1 1.5.2.2 1.5.2.3 1.5.2.4 1.5.2.5 1.5.2.6 1.5.2.7 1.5.2.81.5.2.9 1.5.3.1 1.5.3.2 1.5.3.3 1.5.3.4 1.5.3.5 1.5.3.6 1.5.3.7 1.5.3.81.5.3.9 1.5.4.1 1.5.4.2 1.5.4.3 1.5.4.4 1.5.4.5 1.5.4.6 1.5.4.7 1.5.4.81.5.4.9 1.5.5.1 1.5.5.2 1.5.5.3 1.5.5.4 1.5.5.5 1.5.5.6 1.5.5.7 1.5.5.81.5.5.9 1.5.6.1 1.5.6.2 1.5.6.3 1.5.6.4 1.5.6.5 1.5.6.6 1.5.6.7 1.5.6.81.5.6.9 1.5.7.1 1.5.7.2 1.5.7.3 1.5.7.4 1.5.7.5 1.5.7.6 1.5.7.7 1.5.7.81.5.7.9 1.5.8.1 1.5.8.2 1.5.8.3 1.5.8.4 1.5.8.5 1.5.8.6 1.5.8.7 1.5.8.81.5.8.9 1.5.9.1 1.5.9.2 1.5.9.3 1.5.9.4 1.5.9.5 1.5.9.6 1.5.9.7 1.5.9.81.5.9.9 1.6.1.1 1.6.1.2 1.6.1.3 1.6.1.4 1.6.1.5 1.6.1.6 1.6.1.7 1.6.1.81.6.1.9 1.6.2.1 1.6.2.2 1.6.2.3 1.6.2.4 1.6.2.5 1.6.2.6 1.6.2.7 1.6.2.81.6.2.9 1.6.3.1 1.6.3.2 1.6.3.3 1.6.3.4 1.6.3.5 1.6.3.6 1.6.3.7 1.6.3.81.6.3.9 1.6.4.1 1.6.4.2 1.6.4.3 1.6.4.4 1.6.4.5 1.6.4.6 1.6.4.7 1.6.4.81.6.4.9 1.6.5.1 1.6.5.2 1.6.5.3 1.6.5.4 1.6.5.5 1.6.5.6 1.6.5.7 1.6.5.81.6.5.9 1.6.6.1 1.6.6.2 1.6.6.3 1.6.6.4 1.6.6.5 1.6.6.6 1.6.6.7 1.6.6.81.6.6.9 1.6.7.1 1.6.7.2 1.6.7.3 1.6.7.4 1.6.7.5 1.6.7.6 1.6.7.7 1.6.7.81.6.7.9 1.6.8.1 1.6.8.2 1.6.8.3 1.6.8.4 1.6.8.5 1.6.8.6 1.6.8.7 1.6.8.81.6.8.9 1.6.9.1 1.6.9.2 1.6.9.3 1.6.9.4 1.6.9.5 1.6.9.6 1.6.9.7 1.6.9.81.6.9.9 1.7.1.1 1.7.1.2 1.7.1.3 1.7.1.4 1.7.1.5 1.7.1.6 1.7.1.7 1.7.1.81.7.1.9 1.7.2.1 1.7.2.2 1.7.2.3 1.7.2.4 1.7.2.5 1.7.2.6 1.7.2.7 1.7.2.81.7.2.9 1.7.3.1 1.7.3.2 1.7.3.3 1.7.3.4 1.7.3.5 1.7.3.6 1.7.3.7 1.7.3.81.7.3.9 1.7.4.1 1.7.4.2 1.7.4.3 1.7.4.4 1.7.4.5 1.7.4.6 1.7.4.7 1.7.4.81.7.4.9 1.7.5.1 1.7.5.2 1.7.5.3 1.7.5.4 1.7.5.5 1.7.5.6 1.7.5.7 1.7.5.81.7.5.9 1.7.6.1 1.7.6.2 1.7.6.3 1.7.6.4 1.7.6.5 1.7.6.6 1.7.6.7 1.7.6.81.7.6.9 1.7.7.1 1.7.7.2 1.7.7.3 1.7.7.4 1.7.7.5 1.7.7.6 1.7.7.7 1.7.7.81.7.7.9 1.7.8.1 1.7.8.2 1.7.8.3 1.7.8.4 1.7.8.5 1.7.8.6 1.7.8.7 1.7.8.81.7.8.9 1.7.9.1 1.7.9.2 1.7.9.3 1.7.9.4 1.7.9.5 1.7.9.6 1.7.9.7 1.7.9.81.7.9.9 1.8.1.1 1.8.1.2 1.8.1.3 1.8.1.4 1.8.1.5 1.8.1.6 1.8.1.7 1.8.1.81.8.1.9 1.8.2.1 1.8.2.2 1.8.2.3 1.8.2.4 1.8.2.5 1.8.2.6 1.8.2.7 1.8.2.81.8.2.9 1.8.3.1 1.8.3.2 1.8.3.3 1.8.3.4 1.8.3.5 1.8.3.6 1.8.3.7 1.8.3.81.8.3.9 1.8.4.1 1.8.4.2 1.8.4.3 1.8.4.4 1.8.4.5 1.8.4.6 1.8.4.7 1.8.4.81.8.4.9 1.8.5.1 1.8.5.2 1.8.5.3 1.8.5.4 1.8.5.5 1.8.5.6 1.8.5.7 1.8.5.81.8.5.9 1.8.6.1 1.8.6.2 1.8.6.3 1.8.6.4 1.8.6.5 1.8.6.6 1.8.6.7 1.8.6.81.8.6.9 1.8.7.1 1.8.7.2 1.8.7.3 1.8.7.4 1.8.7.5 1.8.7.6 1.8.7.7 1.8.7.81.8.7.9 1.8.8.1 1.8.8.2 1.8.8.3 1.8.8.4 1.8.8.5 1.8.8.6 1.8.8.7 1.8.8.81.8.8.9 1.8.9.1 1.8.9.2 1.8.9.3 1.8.9.4 1.8.9.5 1.8.9.6 1.8.9.7 1.8.9.81.8.9.9 1.9.1.1 1.9.1.2 1.9.1.3 1.9.1.4 1.9.1.5 1.9.1.6 1.9.1.7 1.9.1.81.9.1.9 1.9.2.1 1.9.2.2 1.9.2.3 1.9.2.4 1.9.2.5 1.9.2.6 1.9.2.7 1.9.2.81.9.2.9 1.9.3.1 1.9.3.2 1.9.3.3 1.9.3.4 1.9.3.5 1.9.3.6 1.9.3.7 1.9.3.81.9.3.9 1.9.4.1 1.9.4.2 1.9.4.3 1.9.4.4 1.9.4.5 1.9.4.6 1.9.4.7 1.9.4.81.9.4.9 1.9.5.1 1.9.5.2 1.9.5.3 1.9.5.4 1.9.5.5 1.9.5.6 1.9.5.7 1.9.5.81.9.5.9 1.9.6.1 1.9.6.2 1.9.6.3 1.9.6.4 1.9.6.5 1.9.6.6 1.9.6.7 1.9.6.81.9.6.9 1.9.7.1 1.9.7.2 1.9.7.3 1.9.7.4 1.9.7.5 1.9.7.6 1.9.7.7 1.9.7.81.9.7.9 1.9.8.1 1.9.8.2 1.9.8.3 1.9.8.4 1.9.8.5 1.9.8.6 1.9.8.7 1.9.8.81.9.8.9 1.9.9.1 1.9.9.2 1.9.9.3 1.9.9.4 1.9.9.5 1.9.9.6 1.9.9.7 1.9.9.81.9.9.9 2.1.1.1 2.1.1.2 2.1.1.3 2.1.1.4 2.1.1.5 2.1.1.6 2.1.1.7 2.1.1.82.1.1.9 2.1.2.1 2.1.2.2 2.1.2.3 2.1.2.4 2.1.2.5 2.1.2.6 2.1.2.7 2.1.2.82.1.2.9 2.1.3.1 2.1.3.2 2.1.3.3 2.1.3.4 2.1.3.5 2.1.3.6 2.1.3.7 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6.5.7.86.5.7.9 6.5.8.1 6.5.8.2 6.5.8.3 6.5.8.4 6.5.8.5 6.5.8.6 6.5.8.7 6.5.8.86.5.8.9 6.5.9.1 6.5.9.2 6.5.9.3 6.5.9.4 6.5.9.5 6.5.9.6 6.5.9.7 6.5.9.86.5.9.9 6.6.1.1 6.6.1.2 6.6.1.3 6.6.1.4 6.6.1.5 6.6.1.6 6.6.1.7 6.6.1.86.6.1.9 6.6.2.1 6.6.2.2 6.6.2.3 6.6.2.4 6.6.2.5 6.6.2.6 6.6.2.7 6.6.2.86.6.2.9 6.6.3.1 6.6.3.2 6.6.3.3 6.6.3.4 6.6.3.5 6.6.3.6 6.6.3.7 6.6.3.86.6.3.9 6.6.4.1 6.6.4.2 6.6.4.3 6.6.4.4 6.6.4.5 6.6.4.6 6.6.4.7 6.6.4.86.6.4.9 6.6.5.1 6.6.5.2 6.6.5.3 6.6.5.4 6.6.5.5 6.6.5.6 6.6.5.7 6.6.5.86.6.5.9 6.6.6.1 6.6.6.2 6.6.6.3 6.6.6.4 6.6.6.5 6.6.6.6 6.6.6.7 6.6.6.86.6.6.9 6.6.7.1 6.6.7.2 6.6.7.3 6.6.7.4 6.6.7.5 6.6.7.6 6.6.7.7 6.6.7.86.6.7.9 6.6.8.1 6.6.8.2 6.6.8.3 6.6.8.4 6.6.8.5 6.6.8.6 6.6.8.7 6.6.8.86.6.8.9 6.6.9.1 6.6.9.2 6.6.9.3 6.6.9.4 6.6.9.5 6.6.9.6 6.6.9.7 6.6.9.86.6.9.9 6.7.1.1 6.7.1.2 6.7.1.3 6.7.1.4 6.7.1.5 6.7.1.6 6.7.1.7 6.7.1.86.7.1.9 6.7.2.1 6.7.2.2 6.7.2.3 6.7.2.4 6.7.2.5 6.7.2.6 6.7.2.7 6.7.2.86.7.2.9 6.7.3.1 6.7.3.2 6.7.3.3 6.7.3.4 6.7.3.5 6.7.3.6 6.7.3.7 6.7.3.86.7.3.9 6.7.4.1 6.7.4.2 6.7.4.3 6.7.4.4 6.7.4.5 6.7.4.6 6.7.4.7 6.7.4.86.7.4.9 6.7.5.1 6.7.5.2 6.7.5.3 6.7.5.4 6.7.5.5 6.7.5.6 6.7.5.7 6.7.5.86.7.5.9 6.7.6.1 6.7.6.2 6.7.6.3 6.7.6.4 6.7.6.5 6.7.6.6 6.7.6.7 6.7.6.86.7.6.9 6.7.7.1 6.7.7.2 6.7.7.3 6.7.7.4 6.7.7.5 6.7.7.6 6.7.7.7 6.7.7.86.7.7.9 6.7.8.1 6.7.8.2 6.7.8.3 6.7.8.4 6.7.8.5 6.7.8.6 6.7.8.7 6.7.8.86.7.8.9 6.7.9.1 6.7.9.2 6.7.9.3 6.7.9.4 6.7.9.5 6.7.9.6 6.7.9.7 6.7.9.86.7.9.9 6.8.1.1 6.8.1.2 6.8.1.3 6.8.1.4 6.8.1.5 6.8.1.6 6.8.1.7 6.8.1.86.8.1.9 6.8.2.1 6.8.2.2 6.8.2.3 6.8.2.4 6.8.2.5 6.8.2.6 6.8.2.7 6.8.2.86.8.2.9 6.8.3.1 6.8.3.2 6.8.3.3 6.8.3.4 6.8.3.5 6.8.3.6 6.8.3.7 6.8.3.86.8.3.9 6.8.4.1 6.8.4.2 6.8.4.3 6.8.4.4 6.8.4.5 6.8.4.6 6.8.4.7 6.8.4.86.8.4.9 6.8.5.1 6.8.5.2 6.8.5.3 6.8.5.4 6.8.5.5 6.8.5.6 6.8.5.7 6.8.5.86.8.5.9 6.8.6.1 6.8.6.2 6.8.6.3 6.8.6.4 6.8.6.5 6.8.6.6 6.8.6.7 6.8.6.86.8.6.9 6.8.7.1 6.8.7.2 6.8.7.3 6.8.7.4 6.8.7.5 6.8.7.6 6.8.7.7 6.8.7.86.8.7.9 6.8.8.1 6.8.8.2 6.8.8.3 6.8.8.4 6.8.8.5 6.8.8.6 6.8.8.7 6.8.8.86.8.8.9 6.8.9.1 6.8.9.2 6.8.9.3 6.8.9.4 6.8.9.5 6.8.9.6 6.8.9.7 6.8.9.86.8.9.9 6.9.1.1 6.9.1.2 6.9.1.3 6.9.1.4 6.9.1.5 6.9.1.6 6.9.1.7 6.9.1.86.9.1.9 6.9.2.1 6.9.2.2 6.9.2.3 6.9.2.4 6.9.2.5 6.9.2.6 6.9.2.7 6.9.2.86.9.2.9 6.9.3.1 6.9.3.2 6.9.3.3 6.9.3.4 6.9.3.5 6.9.3.6 6.9.3.7 6.9.3.86.9.3.9 6.9.4.1 6.9.4.2 6.9.4.3 6.9.4.4 6.9.4.5 6.9.4.6 6.9.4.7 6.9.4.86.9.4.9 6.9.5.1 6.9.5.2 6.9.5.3 6.9.5.4 6.9.5.5 6.9.5.6 6.9.5.7 6.9.5.86.9.5.9 6.9.6.1 6.9.6.2 6.9.6.3 6.9.6.4 6.9.6.5 6.9.6.6 6.9.6.7 6.9.6.86.9.6.9 6.9.7.1 6.9.7.2 6.9.7.3 6.9.7.4 6.9.7.5 6.9.7.6 6.9.7.7 6.9.7.86.9.7.9 6.9.8.1 6.9.8.2 6.9.8.3 6.9.8.4 6.9.8.5 6.9.8.6 6.9.8.7 6.9.8.86.9.8.9 6.9.9.1 6.9.9.2 6.9.9.3 6.9.9.4 6.9.9.5 6.9.9.6 6.9.9.7 6.9.9.86.9.9.9

Additional examples of compounds falling within the scope of Formula Iand Formula II include the following:

The compounds of the present invention incorporate a carbonate group orderivative thereof when X′ is O, S, NR²⁰, or S—O (wherein R²⁰ is H,optionally substituted alkyl, aryl, arylalkyl, C₃₋₆ cycloalkyl, OH, OR²,or O(C═O)R²¹, wherein R²¹ is H, lower alkyl or C₃₋₆ cycloalkyl) attachedto the 2′C and 3′C positions on the ribose sugar as indicated below inthe boxed portion of Formula I:

The presence of the carbonate group leads to surprisingly enhancedproperties of the compounds of the present invention when compared tothe same compounds without a carbonate group at the same location. Thecompounds of the present invention have improved pharmacologicalproperties including one or more of the following: enhanced absorption,increased chemical stability, increased metabolic stability, andincreased liver distribution.

Although not being bound to any particular mechanism, it is believedthat the unexpected improvement in pharmacological properties involvedone or more of the following:

a) enhanced absorption through improvements in the physical propertiesof the nucleoside, including one of more of the following: increasedlipophilicity, decreased solvation, increased solubility, and increaseddissolution in biological fluids;

b) decreased chemical instability that limits oral absorption or firstpass liver exposure. The increased stability results from changes in thephysical properties of the compound, including one or more of thefollowing:

-   -   i) decreased chemical instability in gastrointestinal tract        through decreasing the susceptibility for glycosyl bond cleavage        through changes in preferred conformation and or electronics in        the vicinity of the glycosyl bond;    -   ii) decreased hydrolysis in gastrointestinal tract through        decreased water exposure as a result of increased lipophilicity;

c) decreased metabolic instability that limits oral absorption or firstpass liver exposure. Increased stability results from changes in thephysical properties of the compound that result in the compound beingless susceptible to enzymes that catalyze its metabolism.

Examples of nucleoside and nucleotide degradation that can be affectedby a 2′,3′-cyclic carbonate include one or more of the following:

-   -   i) decreased deamination by enzymes known to catalyze purine or        pyrimidine base deamination. These enzymes limit absorption of        certain nucleosides, especially nucleoside containing adenine-        and cytidine-related analogues. Enzymes known to catalyze        deamination include cytosine deaminase, adenosine deaminase and        adenylate deaminase. One or more of the 2′ and more often 3′        hydroxyls of ribofuranosyl-containing nucleosides and        nucleotides (e.g. with AMP deaminase) interact with protein        residues (cytidine deaminase, Marquez 1984), (adenosine        deaminase, Sharff, 1992). Cyclic carbonates remove both the 2′        and 3′ hydroxyl which are known to aid in catalytic efficiency.    -   ii) decreased glycosyl bond cleavage by nucleosidases. These        enzymes limit absorption by catalyzing the cleavage of the C—N        bond of purine, pyrimidine and other related nucleosides. For        example, purine nucleoside phosphorylase which is sensitive to        modifications at 3′ (Parks et al., 1981). Cyclic carbonates        remove both the 2′ and 3′ hydroxyl which are known to aid in        catalytic efficiency.    -   iii) decreased modification of either or both the 2′ or 3′        hydroxyl by enzymes that catalyze their oxidation to a ketone or        their derivatization to products such as a glucoronide,        sulphate, phosphate, or acylated analogue.

The improved properties of the carbonate compounds of the presentinvention make them particularly useful for the sustained delivery ofnucleoside- and nucleotide-containing compounds. Standard prodrugs, suchas acylated analogues of nucleosides undergo rapid hydrolysis in vivoresulting in the rapid production of the nucleoside. Compounds thatcleave more slowly are useful for sustained delivery of the active drug(nucleoside or phosphorylated metabolites of the nucleoside).

The improved properties of the carbonate compounds of the presentinvention, including increased liver distribution/targeting, also makethem particularly useful for the liver-delivery of nucleoside- andnucleotide-containing compounds. Activation of the prodrug in the livercan result in increased drug levels in the liver and improved efficacy,decreased drug levels outside of the liver and therefore improvedsafety, or both. Prodrugs that are efficiently activated by enzymeswidely distributed throughout the body often result in wide drugdistribution and in the case of nucleosides in a variety of toxicities,including for example neuropathies, myelosuppression, gastrointestinaltoxicity, renal toxicity and cardiovascular toxicity.

The improved properties of the carbonate compounds of the presentinvention make them particularly useful for the treatment of chronicliver diseases, including viral hepatitis, primary liver cancer, cancersthat metastasize to the liver, liver fibrosis, and metabolic diseasesthat involve pathways in the liver that are sensitive to nucleosides andphosphorylated metabolites of nucleosides, including diabetes,hyperlipidemia, obesity and non-alcoholic steatohepatitis.

The compounds of the present invention are also useful for the treatmentof nucleoside and nucleotide responsive diseases including the treatmentof liver diseases responsive to nucleotides which include hepatitis B,hepatitis C, and other viruses that result in viral hepatitis, primaryliver cancer, secondary liver cancer. The compounds of the presentinvention are also useful for the treatment of diseases outside of theliver but responsive to nucleotide analogues, including viralinfections, and cancer. The compounds of the present invention are alsouseful for the treatment of diseases outside the liver that areresponsive to nucleotides and nucleoside analogues which includerespiratory syncytial virus (RSV), herpes simplex type 1 and 2, herpesgenitalis, herpes keratitis, herpes encephalitis, herpes zoster, humanimmunodeficiency virus (HIV), influenza A virus, hantann virus(hemorrhagic fever), human papilloma virus (HPV), measles, fungalinfections, protozoan infections, antiplatelet therapy (P2 receptorantagonists), diabetes (e.g., compounds that bind to the adenosinereceptor, P2 receptor ligands, AMP-activated protein kinase (“AMPK”)activators, cardiovascular disease (e.g., with adenosine based compoundsthat are agonists or antagonists for the adenosine receptor),immunostimulants (e.g., inosine-5′-monophosphate dehydrogenase (“IMPDH”)inhibitors, guanosine-related compounds), inflammation (e.g.,anenosine-related compounds), CNS disorders (sleep, seizure, stroke,pain all of which can be affected by, e.g., adenosine analogues).

Activation of the prodrug compounds of this invention results in theproduction of a nucleoside monophosphate (“NMP”). NMPs are frequentlyfurther phosphorylated inside the hepatocyte to the biologically activenucleoside triphosphate (“NTP”). Drug elimination from the hepatocytetypically entails degradation of phosphorylated metabolites back to aspecies capable of being transported out of the hepatocyte and into theblood for elimination by the kidney or into the bile for biliaryexcretion. Often with nucleoside-based drugs, the phosphorylatedmetabolites are dephosphorylated to the uncharged nucleoside.

Nucleosides that leak back into the systemic circulation result insystemic exposure. If the nucleoside is active systemically, e.g.through entry into virally infected cells and phosphorylation to theactive species, escape of the nucleoside from the liver leads tobiological activity outside of the liver (i.e. extrahepatic tissues,blood cells). In this case, prodrugs of the invention can be effectivefor treating diseases outside of the liver, e.g. viral infections. Sincemany nucleosides exhibit poor oral bioavailability due to breakdown inthe gastrointestinal tract either enzymatically (e.g. deamination byadenosine deaminase) or chemically (e.g. acid instability), the prodrugcan be used for oral drug delivery. Moreover, given that the prodrugs insome cases are broken down slowly relative to e.g. most ester basedprodrugs, the prodrugs could advantageously result in slow, sustainedsystemic release of the nucleoside.

In other cases, however, systemic exposure to the nucleoside can resultin toxicity. This can be minimized by selecting nucleosides that arepreferentially excreted through the bile or nucleosides that are unableto undergo phosphorylation in tissues or nucleosides that undergo rapidintrahepatic metabolism to a biologically inactive metabolite. Someenzymes in the hepatocyte are present that can degrade nucleosides andtherefore minimize exposure (e.g. Phase I and Phase II enzymes). Oneexample is adenosine deaminase, which can deaminate some adenosine-basednucleosides to produce the corresponding inosine analogue. Rapidintracellular deamination of the nucleoside following itsdephosphorylation to the nucleoside limits systemic exposure to thenucleoside and diminishes the risk of toxicity.

Methods described in Examples A-D of the Biological Examples sectionbelow are used to test activation of compounds of this invention.Methods in Example E can be used to evaluate the ability of compounds ofthe invention to generate NTPs.

HCV replication in human liver tissue is evaluated as in Example F.Liver specificity of the prodrugs relative to the nucleosides ismeasured by methods in Example G.

Tissue distribution can be determined according to methods in Example H.Oral bioavailability is determined by methods described in Example I.The susceptibility of nucleoside analogs to metabolism can be determinedas in Example J.

In some aspects of the present invention, the RNA-dependent RNA viralinfection is a positive-sense single-stranded RNA-dependent viralinfection. In other aspects, the positive-sense single-strandedRNA-dependent RNA viral infection is Flaviviridae viral infection orPicornaviridae viral infection. In a subclass of this class, thePicornaviridae viral infection is rhinovirus infection, poliovirusinfection, or hepatitis A virus infection. In a second subclass of thisclass, the Flaviviridae viral infection is selected from the groupconsisting of hepatitis C virus infection, yellow fever virus infection,dengue virus infection, West Nile virus infection, Japanese encephalitisvirus infection, Banzi virus infection, and bovine viral diarrhea virusinfection. In a subclass of this subclass, the Flaviviridae viralinfections hepatitis C virus infection.

In further aspects, compounds of the present invention can be used toenhance the oral bioavailability of the parent drug. In some aspects,compounds of the present invention can be used to enhance the oralbioavailability of the parent drug by at least 5%. In other aspects,compounds of the present invention can be used to enhance the oralbioavailability of the parent drug by at least 10%. In yet otheraspects, oral bioavailability is enhanced by 50% compared to the parentdrug administered orally. In further aspects, the oral bioavailabilityis enhanced by at least 100%.

In some aspects, compounds of the present invention can be used toincrease the therapeutic index of a drug.

In some aspects, compounds of the present invention can be used tobypass drug resistance.

In other aspects, compounds of the present invention can be used totreat cancer.

Thus, the present invention provides methods for inhibiting viralreplication comprising the step of administering to a patient atherapeutically effective amount of a compound of the present invention,or a solvate, hydrate, prodrug, or pharmaceutically acceptable saltthereof.

The present invention also provides methods for inhibiting RNA-dependentRNA viral replication comprising the step of administering to a patienta therapeutically effective amount of a compound of the presentinvention, or a solvate, hydrate, prodrug, or pharmaceuticallyacceptable salt thereof.

The present invention further provides methods for inhibiting HCVreplication comprising the step of administering to a patient atherapeutically effective amount of a compound of the present invention,or a solvate, hydrate, prodrug, or pharmaceutically acceptable saltthereof.

The present invention also provides methods for treating viralinfections comprising the step of administering to a patient atherapeutically effective amount of a compound of the present invention,or a solvate, hydrate, prodrug, or pharmaceutically acceptable saltthereof.

The present invention also provides methods for treating viralinfections of the liver comprising the step of administering to apatient a therapeutically effective amount of a compound of the presentinvention, or a solvate, hydrate, prodrug, or pharmaceuticallyacceptable salt thereof.

The present invention also provides methods for treating RNA-dependentRNA viral infection comprising the step of administering to a patient atherapeutically effective amount of a compound of the present invention,or a solvate, hydrate, prodrug, or pharmaceutically acceptable saltthereof.

The present invention also provides methods for treating hepatitis Bvirus (HBV) or hepatitis C virus (HCV) infection comprising the step ofadministering to a patient a therapeutically effective amount of acompound of the present invention, or a solvate, hydrate, prodrug, orpharmaceutically acceptable salt thereof.

The present invention also provides methods for treating chronic liverdiseases, including viral hepatitis, primary liver cancer, cancers thatmetastasize to the liver, liver fibrosis, and metabolic diseases thatinvolve pathways in the liver that are sensitive to nucleosides andphosphorylated metabolites of nucleosides, including diabetes,hyperlipidemia, obesity and non-alcoholic steatohepatitis comprising thestep of administering to a patient a therapeutically effective amount ofa compound of the present invention, or a solvate, hydrate, prodrug, orpharmaceutically acceptable salt thereof.

The present invention also provides methods for treating a plateletdisorder, or diabetes comprising the step of administering to a patienta therapeutically effective amount of a compound of the presentinvention that is a P2 receptor antagonist, or a solvate, hydrate,prodrug, or pharmaceutically acceptable salt thereof.

The present invention also provides methods for treating diabetescomprising the step of administering to a patient a therapeuticallyeffective amount of a compound of the present invention that is an AMPKactivator, or a solvate, hydrate, prodrug, or pharmaceuticallyacceptable salt thereof.

The present invention also provides methods for treating diabetes orcardiovascular disease comprising the step of administering to a patienta therapeutically effective amount of a compound of the presentinvention that binds an adenosine receptor, or a solvate, hydrate,prodrug, or pharmaceutically acceptable salt thereof.

The present invention also provides methods for treating animmunodeficiency disease comprising the step of administering to apatient a therapeutically effective amount of a compound of the presentinvention that acts as an immunostimulant inhibiting IMPDH, or asolvate, hydrate, prodrug, or pharmaceutically acceptable salt thereof.

The present invention also provides methods for treating inflammation ora CNS disorder comprising the step of administering to a patient atherapeutically effective amount of a compound of the present inventionthat acts as an adenosine analogue, or a solvate, hydrate, prodrug, orpharmaceutically acceptable salt thereof.

Formulations

The compounds of the present invention are administered in a total dailydose of 0.01 to 1000 mg/kg. In some aspects of the invention, the rangeis about 0.1 mg/kg to about 100 mg/kg. In other aspects, the range is0.5 to 20 mg/kg. The dose may be administered in as many divided dosesas is convenient.

Compounds of this invention when used in combination with otherantiviral agents may be administered as a daily dose or an appropriatefraction of the daily dose (e.g., bid). Administration of the prodrugcompound may occur at or near the time in which the other antiviral isadministered or at a different time. The compounds of this invention maybe used in a multidrug regimen, also known as combination or ‘cocktail’therapy, wherein, multiple agents may be administered together, may beadministered separately at the same time or at different intervals, oradministered sequentially. The compounds of this invention may beadministered after a course of treatment by another agent, during acourse of therapy with another agent, administered as part of atherapeutic regimen, or can be administered prior to therapy by anotheragent in a treatment program.

For the purposes of this invention, the compounds may be administered bya variety of means including orally, parenterally, by inhalation spray,topically, or rectally in formulations containing pharmaceuticallyacceptable carriers, adjuvants and vehicles. The term parenteral as usedhere includes subcutaneous, intravenous, intramuscular, andintra-arterial injections with a variety of infusion techniques.Intra-arterial and intravenous injection as used herein includesadministration through catheters.

Intravenous administration is generally preferred.

Pharmaceutically acceptable salts include acetate, adipate, besylate,bromide, camsylate, chloride, citrate, edisylate, estolate, fumarate,gluceptate, gluconate, glucuronate, hippurate, hyclate, hydrobromide,hydrochloride, iodide, isethionate, lactate, lactobionate, maleate,mesylate, methylbromide, methylsulfate, napsylate, nitrate, oleate,palmoate, phosphate, polygalacturonate, stearate, succinate, sulfate,sulfosalicylate, tannate, tartrate, terphthalate, tosylate, andtriethiodide.

Pharmaceutical compositions containing the active ingredient may be inany form suitable for the intended method of administration. When usedfor oral use for example, tablets, troches, lozenges, aqueous or oilsuspensions, dispersible powders or granules, emulsions, hard or softcapsules, syrups or elixirs may be prepared. Compositions intended fororal use may be prepared according to any method known to the art forthe manufacture of pharmaceutical compositions and such compositions maycontain one or more agents including sweetening agents, flavoringagents, coloring agents and preserving agents, in order to provide apalatable preparation. Tablets containing the active ingredient inadmixture with non-toxic pharmaceutically acceptable excipients whichare suitable for manufacture of tablets are acceptable. These excipientsmay be, for example, inert diluents, such as calcium or sodiumcarbonate, lactose, calcium or sodium phosphate; granulating anddisintegrating agents, such as maize starch, or alginic acid; bindingagents, such as starch, gelatin or acacia; and lubricating agents, suchas magnesium stearate, stearic acid or talc. Tablets may be uncoated orcan be coated by known techniques including microencapsulation to delaydisintegration and adsorption in the gastrointestinal tract and therebyprovide a sustained action over a longer period. For example, a timedelay material such as glyceryl monostearate or glyceryl distearatealone or with a wax may be employed.

Formulations for oral use may be also presented as hard gelatin capsuleswhere the active ingredient is mixed with an inert solid diluent, forexample calcium phosphate or kaolin, or as soft gelatin capsules whereinthe active ingredient is mixed with water or an oil medium, such aspeanut oil, liquid paraffin or olive oil.

Aqueous suspensions of the invention contain the active materials inadmixture with excipients suitable for the manufacture of aqueoussuspensions. Such excipients include a suspending agent, such as sodiumcarboxymethylcellulose, methylcellulose, ethylcellulose,hydroxypropylcellulose, hydroxypropyl methylcellulose, sodium alginate,polyvinylpyrrolidone, gum tragacanth and gum acacia, and dispersing orwetting agents such as a naturally occurring phosphatide (e.g.,lecithin), a condensation product of an alkylene oxide with a fatty acid(e.g., polyoxyethylene stearate), a condensation product of ethyleneoxide with a long chain aliphatic alcohol (e.g.,heptadecaethyleneoxycetanol), a condensation product of ethylene oxidewith a partial ester derived from a fatty acid and a hexitol anhydride(e.g., polyoxyethylene sorbitan monooleate). The aqueous suspension mayalso contain one or more preservatives such as ethyl or n-propylp-hydroxy-benzoate, one or more coloring agents, one or more flavoringagents and one or more sweetening agents, such as sucrose or saccharin.

Oil suspensions may be formulated by suspending the active ingredient ina vegetable oil, such as arachid oil, olive oil, sesame oil or coconutoil, or in a mineral oil such as liquid paraffin. The oral suspensionsmay contain a thickening agent, such as beeswax, hard paraffin or cetylalcohol. Sweetening agents, such as those set forth above, and flavoringagents may be added to provide a palatable oral preparation. Thesecompositions may be preserved by the addition of an antioxidant such asascorbic acid.

Dispersible powders and granules of the invention suitable forpreparation of an aqueous suspension by the addition of water providethe active ingredient in admixture with a dispersing or wetting agent, asuspending agent, and one or more preservatives. Suitable dispersing orwetting agents and suspending agents are exemplified by those disclosedabove. Additional excipients, for example sweetening, flavoring andcoloring agents, may also be present.

The pharmaceutical compositions of the invention may also be in the formof oil-in-water emulsions. The oily phase may be a vegetable oil, suchas olive oil or arachid oil, a mineral oil, such as liquid paraffin, ora mixture of these. Suitable emulsifying agents includenaturally-occurring gums, such as gum acacia and gum tragacanth,naturally occurring phosphatides, such as soybean lecithin, esters orpartial esters derived from fatty acids and hexitol anhydrides, such assorbitan monooleate, and condensation products of these partial esterswith ethylene oxide, such as polyoxyethylene sorbitan monooleate. Theemulsion can also contain sweetening and flavoring agents.

Syrups and elixirs may be formulated with sweetening agents, such asglycerol, sorbitol or sucrose. Such formulations may also contain ademulcent, a preservative, a flavoring or a coloring agent.

The pharmaceutical compositions of the invention may be in the form of asterile injectable preparation, such as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according tothe known art using those suitable dispersing or wetting agents andsuspending agents which have been mentioned above. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally acceptable diluent or solvent,such as a solution in 1,3-butane-diol or prepared as a lyophilizedpowder. Among the acceptable vehicles and solvents that may be employedare water, Ringer's solution and isotonic sodium chloride solution. Inaddition, sterile fixed oils may conventionally be employed as a solventor suspending medium. For this purpose any bland fixed oil may beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid may likewise be used in the preparation ofinjectables.

The amount of active ingredient that may be combined with the carriermaterial to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. For example, atime-release formulation intended for oral administration to humans maycontain 20 to 2000 μmol (approximately 10 to 1000 mg) of active materialcompounded with an appropriate and convenient amount of carrier materialwhich may vary from about 5 to about 95% of the total compositions. Itis preferred that the pharmaceutical composition be prepared whichprovides easily measurable amounts for administration. For example, anaqueous solution intended for intravenous infusion should contain fromabout 0.05 to about 50 μmol (approximately 0.025 to 25 mg) of the activeingredient per milliliter of solution in order that infusion of asuitable volume at a rate of about 30 mL/h can occur.

As noted above, formulations of the present invention suitable for oraladministration may be presented as discrete units such as capsules,cachets or tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous or non-aqueous liquid; or as an oil-in-water liquid emulsion ora water-in-oil liquid emulsion. The active ingredient may also beadministered as a bolus, electuary or paste.

A tablet may be made by compression or molding, optionally with one ormore accessory ingredients. Compressed tablets may be prepared bycompressing in a suitable machine the active ingredient in a freeflowing form such as a powder or granules, optionally mixed with abinder (e.g., povidone, gelatin, hydroxypropylmethyl cellulose),lubricant, inert diluent, preservative, disintegrant (e.g., sodiumstarch glycolate, cross-linked povidone, cross-linked sodiumcarboxymethyl cellulose) surface active or dispersing agent. Moldedtablets may be made by molding in a suitable machine a mixture of thepowdered compound moistened with an inert liquid diluent. The tabletsmay optionally be coated or scored and may be formulated so as toprovide slow or controlled release of the active ingredient thereinusing, for example, hydroxypropyl methylcellulose in varying proportionsto provide the desired release profile. Tablets may optionally beprovided with an enteric coating, to provide release in parts of the gutother than the stomach. This is particularly advantageous with thecompounds of Formula I when such compounds are susceptible to acidhydrolysis.

Formulations suitable for topical administration in the mouth includelozenges comprising the active ingredient in a flavored base, usuallysucrose and acacia or tragacanth; pastilles comprising the activeingredient in an inert base such as gelatin and glycerin, or sucrose andacacia; and mouthwashes comprising the active ingredient in a suitableliquid carrier.

Formulations for rectal administration may be presented as a suppositorywith a suitable base comprising for example cocoa butter or asalicylate.

Formulations suitable for vaginal administration may be presented aspessaries, tampons, creams, gels, pastes, foams or spray formulationscontaining in addition to the active ingredient such carriers as areknown in the art to be appropriate.

Formulations suitable for parenteral administration include aqueous andnon-aqueous isotonic sterile injection solutions which may containantioxidants, buffers, bacteriostats and solutes which render theformulation isotonic with the blood of the intended recipient; andaqueous and non-aqueous sterile suspensions which may include suspendingagents and thickening agents. The formulations may be presented inunit-dose or multi-dose sealed containers, for example, ampoules andvials, and may be stored in a freeze-dried (lyophilized) conditionrequiring only the addition of the sterile liquid carrier, for examplewater for injections, immediately prior to use.

Injection solutions and suspensions may be prepared from sterilepowders, granules and tablets of the kind previously described.

Formulations suitable for parenteral administration may be administeredin a continuous infusion manner via an indwelling pump or via a hospitalbag. Continuous infusion includes the infusion by an external pump. Theinfusions may be done through a Hickman or PICC or any other suitablemeans of administering a formulation either parenterally or i.v.

Preferred unit dosage formulations are those containing a daily dose orunit, daily sub-dose, or an appropriate fraction thereof, of a drug.

It will be understood, however, that the specific dose level for anyparticular patient will depend on a variety of factors including theactivity of the specific compound employed; the age, body weight,general health, sex and diet of the individual being treated; the timeand route of administration; the rate of excretion; other drugs whichhave previously been administered; and the severity of the particulardisease undergoing therapy, as is well understood by those skilled inthe art.

Another aspect of the present invention is concerned with a method ofinhibiting HCV replication or treating HCV infection with a compound ofthe present invention in combination with one or more agents useful fortreating HCV infection. Such agents active against HCV include, but arenot limited to, ribavirin, levovirin, viramidine, thymosin alpha-1,interferon-β, interferon-α, pegylated interferon-α (peginterferon-α), acombination of interferon-α and ribavirin, a combination ofpeginterferon-α and ribavirin, a combination of interferon-α andlevovirin, and a combination of peginterferon-α and levovirin.Interferon-α includes, but is not limited to, recombinant interferon-α2a(such as Roferon interferon available from Hoffmann-LaRoche, Nutley,N.J.), pegylated interferon-α2a (Pegasys™), interferon-α2b (such asIntron-A interferon available from Schering Corp., Kenilworth, N.J.),pegylated interferon-α2b (Peglntron™), a recombinant consensusinterferon (such as interferon alphacon-1), and a purified interferon-αproduct. Amgen's recombinant consensus interferon has the brand nameInfergen®. Levovirin is the L-enantiomer of ribavirin which has shownimmunomodulatory activity similar to ribavirin. Viramidine is aliver-targeting prodrug analog of ribavirin disclosed in InternationalPubl. No. WO 01/60379 (assigned to ICN Pharmaceuticals). In accordancewith this method of the present invention, the individual components ofthe combination can be administered separately at different times duringthe course of therapy or concurrently in divided or single combinationforms. The instant invention is therefore to be understood as embracingall such regimes of simultaneous or alternating treatment, and the term“administering” is to be interpreted accordingly. It will be understoodthat the scope of combinations of the compounds of this invention withother agents useful for treating HCV infection includes in principle anycombination with any pharmaceutical composition for treating HCVinfection. When a compound of the present invention or apharmaceutically acceptable salt thereof is used in combination with asecond therapeutic agent active against HCV, the dose of each compoundmay be either the same as or different from the dose when the compoundis used alone.

Also included within the scope of the invention is a pharmaceuticalcomposition comprising a compound of Formula I, solvate, hydrate,prodrug or pharmaceutically acceptable salt thereof, and at least oneagent useful for treating a viral infection, particularly an HCVinfection.

For the treatment of HCV infection, the compounds of the presentinvention may also be administered in combination with an agent that isan inhibitor of HCV NS3 serine protease. HCV NS3 serine protease is anessential viral enzyme and has been described to be an excellent targetfor inhibition of HCV replication. Both substrate and non-substratebased inhibitors of HCV NS3 protease inhibitors are disclosed inInternational Publ. Nos. WO 98/22496, WO 98/46630, WO 99/07733, WO99/07734, WO 99/38888, WO 99/50230, WO 99/64442, WO 00/09543, WO00/59929, WO 02/48116, WO 02/48172; in GB-2337262; and in U.S. Pat. Nos.6,323,180 and 6,410,531. Specific embodiments of NS3 protease inhibitorsfor combination with the compounds of the present invention are BILN2061 (Boehringer Ingelheim) and VX-950/LY-570310. HCV NS3 protease as atarget for the development of inhibitors of HCV replication and for thetreatment of HCV infection is discussed in Dymock, B. W., “Emergingtherapies for hepatitis C virus infection,” Emerging Drugs 6:13-42(2001).

Ribavirin, levovirin, and viramidine may exert their anti-HCV effects bymodulating intracellular pools of guanine nucleotides via inhibition ofthe intracellular enzyme inosine monophosphate dehydrogenase (“IMPDH”).IMPDH is the rate-limiting enzyme on the biosynthetic route in de novoguanine nucleotide biosynthesis. Ribavirin is readily phosphorylatedintracellularly and the monophosphate derivative is an inhibitor ofIMPDH. Thus, inhibition of IMPDH represents another useful target forthe discovery of inhibitors of HCV replication. Therefore, the compoundsof the present invention may also be administered in combination with aninhibitor of IMPDH, such as VX-497 (merimepodib), which is disclosed inInternational Publ. Nos. WO 97/41211 and WO 01/00622 (assigned toVertex); another IMPDH inhibitor, such as that disclosed inInternational Publ. No. WO 00/25780 (assigned to Bristol-Myers Squibb);or mycophenolate mofetil (see Allison, A. C. and Eugui, E. M., AgentsAction 44 (Suppl):165 (1993)).

For the treatment of HCV infection, the compounds of the presentinvention may also be administered in combination with the antiviralagent amantadine (1-aminoadarnantane) and its hydrochloride salt (for acomprehensive description of this agent, see Kirschbaum, J., Anal.Profiles Drug Subs. 12:1-36 (1983)).

The compounds of the present invention may also be combined for thetreatment of HCV infection with antiviral 1-C, 2′-C—, or 3′-C-branchedribonucleosides disclosed in R. E. Harry-O'kuru, et al., J. Org. Chem.62:1754-1759 (1997); M. S. Wolfe, et al., Tetrahedron Lett. 36:7611-7614(1995); U.S. Pat. No. 3,480,613 (Nov. 25, 1969); International Publ. No.WO 01/90121 (29 Nov. 2001); International Publ. No. WO 01/92282 (6 Dec.2001); and International Publ. No. WO 02/32920 (25 Apr. 2002); thecontents of each of which are incorporated by reference in theirentirety. Such branched ribonucleosides include, but are not limited to,2′-C-methylcytidine, 2′-C-methyluridine, 2′-C-methyladenosine,2′-C-methylguanosine, and9-(2-C-methyl-β-D-ribofuranosyl)-2,6-diaminopurine, and prodrugsthereof.

The compounds of the present invention may also be combined for thetreatment of HCV infection with other nucleosides having anti-HCVproperties, such as those disclosed in International Publ. No. WO02/51425 (4 Jul. 2002), assigned to Mitsubishi Pharma Corp.;International Publ. Nos.WO 01/79246, WO 02/32920 (25 Apr. 2002), and WO02/48165 (20 Jun. 2002), assigned to Pharmasset, Ltd.; InternationalPubl. No. WO 01/68663 (20 Sep. 2001), assigned to ICN Pharmaceuticals;International Publ. No. WO 99/43691 (2 Sep. 1999); International Publ.No. WO 02/18404 (7 Mar. 2002), assigned to Hoffmann-LaRoche; U.S.2002/0019363 (14 Feb. 2002); International Publ. No. WO 02/057287 (25Jul. 2002), assigned to Merck & Co. and Isis Pharmaceuticals; andInternational Publ. No. WO 02/057425 (25 Jul. 2002), assigned to Merck &Co. and Isis Pharmaceuticals.

The compounds of the present invention may also be combined for thetreatment of HCV infection with non-nucleoside inhibitors of HCVpolymerase such as those disclosed in International Publ. No. WO01/77091 (18 Oct. 2001), assigned to Tularik, Inc.; International Publ.No. WO 01/47883 (5 Jul. 2001), assigned to Japan Tobacco, Inc.;International Publ. No. WO 02/04425 (17 Jan. 2002), assigned toBoehringer Ingelheim; International Publ. No. WO 02/06246 (24 Jan.2002), assigned to Istituto di Ricerche di Biologia Moleculare P.Angeletti S. P. A.; and International Publ. No. WO 02/20497 (3 Mar.2002). International Publ. No. WO 01/47883 discloses a large number ofbenzimidazole derivatives, such as JTK-003, which is claimed to be anorally active inhibitor of NS5B that is currently undergoing clinicalevaluation.

Synthesis of Nucleoside Compound Derivatives

The following description provides procedures for synthesizing2′,3′-cyclic carbonate NMP prodrugs of the present invention and isorganized into three sections: (1) synthesis of 2′,3′-carbonates, (2)synthesis of phosphorylation precursors, and (3) synthesis of NMPprodrugs.

Synthesis of 2′,3′-cyclic Carbonate of Nucleoside Derivatives:

Synthesis of 2′,3′-carbonate of nucleoside derivatives of formula I maybe organized into two following sections: (i) synthesis of2′,3′-carbonate of nucleoside analogs; and (ii) synthesis of2′,3′-carbonate of nucleotide analogs.

(i) Synthesis of 2′,3′ Cyclic Carbonate of Nucleoside Analogs:

Synthesis of 2′,3′-carbonate of nucleoside analogs may be achieved by avariety of known methods (Greene T. W., Protective Groups in OrganicChemistry, John Wiley & Sons, New York (1999)). Following are twogeneral routes wherein path A is an approach via carbonylation of2′,3′-vicinal diols of 5′-hydroxy protected nucleosides and path B is anapproach wherein direct carbonylation is attained on unprotectednucleosides.

Synthesis via path A includes carbonylation of nucleosides with masked5′-hydroxy group. Protection of the 5′-hydroxy group may be attained byacid labile functionality such as silyl or trityl groups. Thesenucleoside derivatives containing a protected 5′-hydroxy then undergocarbonylation via a range of reagents, such as N,N′-carbonyl diimidazole(Kutney et al., Synth. Commun. 5:47 (1975)) or p-nitrophenylchloroformate (Cook et al., J. Org. Chem. 33:3589 (1968)) under mildconditions. Such methods are applicable to a variety of nucleosides withdiverse sugar as well as base substitutions. The final step of thesequence (path A) involves removal of the 5′-protective group undermildly acidic or neutral reaction conditions.

Synthesis of 2′,3′-carbonate of ribonucleoside derivatives via directcarbonylation as shown in path B can be attained without a protectivegroup at the 5′-position. Thermal reaction of diaryl carbonates uponreaction with nucleosides in polar solvents such as DMF orN-methylpyrrolidine gives rapid conversion of nucleosides to the desired2′,3′-carbonate derivatives (Hampton et al., Biochemistry 5:2076(1966)). Such reactions with diphenyl carbonate were found to beaccelerated by microwave mediated thermal conditions. Directcarbonylation can also be achieved with p-nitrophenylchloroformate inthe case of uridine nucleoside analogs (Letsinger et al., J Org. Chem.32:296 (1967)).

(ii) Synthesis of 2′,3′-carbonate of Nucleotide Analogs.

2′,3′-Carbonate containing nucleotide analogs may be prepared via twodifferent protocols. These compounds can be made starting from2′,3′-carbonate derivative of nucleosides as shown via path A or frompath B from phosphorylated nucleoside analogs. As in path A,2′,3′-carbonate derivative of nucleosides can be phosphorylated to givenucleoside monophosphate (NMP) prodrugs via phosphorylation utilizingP(EI) or P(V) intermediates (Maclman et al., Ann. Rep. Med. Chem. 39:305(2004)). Alternatively, such prodrugs may be prepared via carbonylationas shown in path B following conditions described in the earliersection.

Synthesis of 2′3′-thionocarbonate Prodrugs:

2′,3′-Thionocarbonate containing compounds of the present invention(compounds where X′ is S) can be made by the procedures described above,by replacing carbonylation reaction with thionocarbonate formation fromthe earlier synthetic sequences. Several well-known methods areavailable for such transformation. For example, thionocarbonateformation can be achieved by treatment of vicinal diols of nucleosidederivatives by 1,1′-thiocarbonyldiimidazole (Yu et al., Org. Lett.4:1919 (2002)) or by 1,1′-thiocarbonyldi-2(1H)-pyridone (Kim et al., J.Org. Chem. 51:2615 (1986)). Reactions of phenyl chlorothionoformate(Halila et al., Carbohydrate Res. 337:69 (2002)), or thiophosgene (He etal., J. Org. Chem. 65:7627 (2000)) are also known to convert vicinal1,2-diols to corresponding thionocarbonates. Such mild reactionconditions can be utilized to form the 2′,3′-thionocarbonatefunctionality at any desired stage of prodrug synthesis.

Synthesis of Phosphorylation Precursors:

Synthesis of phosphorylation precursors is attained in two stages: 1.Synthesis of 1,3-diols; and 2. Synthesis of phosphorylation precursor.

Synthesis of 1,3-Diols:

A variety of synthetic methods are known to prepare the following typesof 1,3-diols: a) 1-substituted; b) 2-substituted; and c) 1,2- or1,3-annulated in their racemic or enantioenriched form. The V, W, Zgroups of Z″ of Formula II (i.e., V, W, and Z groups of Formula II) canbe introduced or modified either during synthesis of diols or after thesynthesis of prodrugs.

Synthesis of 1-(aryl)-Propane-1,3-Diols:

The suitable methods to prepare 1,3-diols are divided into two types asfollowing: 1) synthesis of racemic 1-(aryl)-propane-1,3-diols; and 2)synthesis of enantioenriched 1-(aryl)-propane-1,3-diols.

Synthesis of Racemic 1-(aryl)-Propane-1,3-Diol:

1,3-Dihydroxy compounds can be synthesized by several well-known methodsfrom the literature. Substituted aromatic aldehydes are utilized tosynthesize racemic 1-(aryl)propane-1,3-diols via addition of lithiumenolate of alkyl acetate followed by ester reduction (path A) (Turner,J. Org. Chem. 55:4744 (1990)). Alternatively, aryl lithium or arylGrignard additions to 1-hydroxy propan-3-al also give1-(arylsubstituted)propane-1,3-diols (path B). This method will enableconversion of various substituted aryl halides to1-(arylsubstituted)-1,3-propane diols (Coppi, et al., J. Org. Chem.53:911 (1988)). Aryl halides can also be used to synthesize1-substituted propane diols by Heck coupling of 1,3-diox-4-ene followedby reduction and hydrolysis (Sakamoto, et al., Tetrahedron Lett. 33:6845(1992)). Pyridyl-, quinolyl-, isoquinolyl-propan-3-ol derivatives can behydroxylated to 1-substituted-1,3-diols by N-oxide formation followed byrearrangement in the presence of acetic anhydride (path C) (Yamamoto, etal., Tetrahedron 37:1871 (1981)). A variety of aromatic aldehydes canalso be converted to 1-substituted-1,3-diols by vinyl lithium or vinylGrignard addition followed by hydroboration reaction (path D).

Synthesis of Enantioenricbed 1-(aryl)-Propane-1,3-Diol:

A variety of known methods for resolution of secondary alcohols viachemical or enzymatic agents may be utilized for preparation of diolenantiomers (Harada, et al., Tetrahedron Lett. 28:4843 (1987)).Transition metal catalyzed hydrogenation of substituted3-aryl-3-oxo-propionic acids or esters is an efficient method to prepareR- or S-isomers of beta hydroxy acids or esters in high enantiomericpurity (Comprehensive Asymmetric Catalysis, Jacobsen, E. N., et al.,eds., Springer (1999); Noyori, R., Asymmetric Catalysis in OrganicSynthesis, John Wiley (1994)). These beta hydroxy acid or ester productscan be further reduced to give required 1-(aryl)-propane-1,3-diols inhigh enantiomeric excess (ee). (path A). The β-keto acid or estersubstrates for high pressure hydrogenation or hydrogen transferreactions may be prepared by a variety of methods such as condensationof acetophenone with dimethylcarbonate in the presence of a base (Chu,et al., J. Het Chem. 22:1033 (1985)), by ester condensation (Turner, etal., J. Org. Chem. 54:4229 (1989)) or from aryl halides (Kobayashi, etal., Tetrahedron Lett. 27:4745 (1986)). Alternatively, 1,3-diols of highenantiomeric purity can be obtained by enantioselective borane reductionof p-hydroxyethyl aryl ketone derivatives or β-keto acid derivatives(path B) (Ramachandran, et al., Tetrahedron Lett. 38:761 (1997)). Inanother method, commercially available cinnamyl alcohols may beconverted to epoxy alcohols under catalytic asymmetric epoxidationconditions. These epoxy alcohols are reduced by Red-Al to result in1,3-diols with high ee's (path C) (Gao, et al., J. Org. Chem. 53:4081(1980)). Enantioselective aldol condensation is another well-describedmethod for synthesis of 1,3-oxygenated functionality with high ee'sstarting from aromatic aldehydes. (path D) (Mukaiyama, Org. React.28:203 (1982)).

Synthesis of 2-Substituted 1,3-Diols:

Various 2-substituted-1,3-diols can be made from commercially available2-(hydroxymethyl)-1,3-propane-diol. Pentaerythritol can be converted totriol via decarboxylation of diacid followed by reduction (path a)(Werle, et al., Liebigs. Ann. Chem. 944 (1986)) or diol-monocarboxylicacid derivatives can also be obtained by decarboxylation under knownconditions (Iwata, et. al., Tetrahedron Lett. 28:3131 (1987)).Nitrotriol is also known to give triol by reductive elimination (path b)(Latour, et. al., Synthesis 8:742 (1987)). The triol can be derivatizedby mono acylation or carbonate formation by treatment with alkanoylchloride, or alkylchloroformate (path d) (Greene and Wuts, Protectivegroups in organic synthesis, John Wiley, New York (1990)). Arylsubstitution can be affected by oxidation to aldehyde and aryl Grignardadditions (path c). Aldehydes can also be converted to substitutedamines by reductive amination reaction (path e).

Synthesis of cyclic-1,3-diols:

Compounds of Formula II where V-Z or V-W are fused by four carbons aremade from cyclohexane diol derivatives. Commercially available cis,cis-1,3,5-cyclohexane-triol can be used as is or modified as describedin case of 2-substituted propan-1,3-diols to give various analogues.These modifications can either be made before or after ester formation.Various 1,3-cyclohexane-diols can be made by Diels-Alder methodologyusing pyrone as diene (Posner, et. al., Tetrahedron Lett. 32:5295(1991)). Cyclohexanediol derivatives are also made by nitrileoxide-olefin additions (Curran, et. al., J. Am. Chem. Soc. 107:6023(1985)). Alternatively, cyclohexyl precursors are also made fromcommercially available quinic acid (Rao, et. al., Tetrahedron Lett.32:547 (1991).)

Synthesis of Substituted 1,3-hydroxyamines and 1,3-diamines:

A large number of synthetic methods are available for the preparation ofsubstituted 1,3-hydroxyamines and 1,3-diamines due to the ubiquitousnature of these functionalities in naturally occurring compounds.Following are some of these methods organized into: 1. synthesis ofsubstituted 1,3-hydroxy amines; 2. synthesis of substituted 1,3-diaminesand 3. synthesis of chiral substituted 1,3-hydroxyamines and1,3-diamines.

Synthesis of Substituted 1,3-hydroxy Amines:

1,3-Diols described in the earlier section can be converted selectivelyto either hydroxy amines or to corresponding diamines by convertinghydroxy functionality to a leaving group and treating with anhydrousammonia or required primary or secondary amines (Corey, et al.,Tetrahedron Lett., 1989, 30, 5207: Gao, et al., J. Org. Chem. 53:4081(1988)). A similar transformation may also be achieved directly fromalcohols under Mitsunobu type of reaction conditions (Hughes, D. L.,Org. React. 42 (1992)).

A general synthetic procedure for 3-aryl-3-hydroxy-propan-1-amine typeof prodrug moiety involves aldol type condensation of aryl esters withalkyl nitrites followed by reduction of resulting substitutedbenzoylacetonitrile (Shih et al., Heterocycles 24:1599 (1986)). Theprocedure can also be adapted for formation of 2-substitutedaminopropanols by using substituted alkylnitrile. In another approach,3-aryl-3-amino-propan-1-ol type of prodrug groups are synthesized fromaryl aldehydes by condensation of malonic acid in presence of ammoniumacetate followed by reduction of resulting substituted β-amino acids.Both these methods enable to introduce wide variety of substitution ofaryl group (Shih, et al., Heterocycles. 9:1277 (1978)). In an alternateapproach, P-substituted organolithium compounds of 1-amino-1-aryl ethyldianion generated from styrene type of compounds undergo addition withcarbonyl compounds to give variety of W, W′ substitution by variation ofthe carbonyl compounds (Barluenga, et al., J. Org. Chem. 44:4798(1979)).

Synthesis of Substituted 1,3-diamines:

Substituted 1,3-diamines are synthesized starting from a variety ofsubstrates. Arylglutaronitriles can be transformed to 1-substituteddiamines by hydrolysis to amide and Hofmann rearrangement conditions(Bertochio, et al., Bull. Soc. Chim. Fr. 1809 (1962)). Whereas,malononitrile substitution will enable variety of Z substitution byelectrophile introduction followed by hydride reduction to correspondingdiamines. In another approach, cinnamaldehydes react with hydrazines orsubstituted hydrazines to give corresponding pyrazolines which uponcatalytic hydrogenation result in substituted 1,3-diamines (Weinhardt,et al., J. Med. Chem. 28:694 (1985)). High trans-diastereoselectivity of1,3-substitution is also attainable by aryl Grignard addition on topyrazolines followed by reduction (Alexakis, et al., J. Org. Chem.576:4563 (1992)). 1-Aryl-1,3-diaminopropanes are also prepared bydiborane reduction of 3-amino-3-arylacrylonitriles which in turn aremade from nitrile substituted aromatic compounds (Domow, et al., ChemBer. 82:254 (1949)). Reduction of 1,3-diimines obtained fromcorresponding 1,3-carbonyl compounds are another source of 1,3-diamineprodrug moiety which allows a wide variety of activating groups V and/orZ (Barluenga, et al., J. Org. Chem. 48:2255 (1983)).

Synthesis of Chiral Substituted 1,3-hydroxyamines and 1,3-diamines:

Enantiomerically pure 3-aryl-3-hydroxypropan-1-amines are synthesized byCBS enantioselective catalytic reaction of β-chloropropiophenonefollowed by displacement of halo group to make secondary or primaryamines as required (Corey, et al., Tetrahedron Lett. 30:5207 (1989)).Chiral 3-aryl-3-amino propan-1-ol type of prodrug moiety may be obtainedby 1,3-dipolar addition of chirally pure olefin and substituted nitroneof arylaldehyde followed by reduction of resulting isoxazolidine(Koizumi, et al., J. Org. Chem. 47:4005 (1982)). Chiral induction in1,3-polar additions to form substituted isoxazolidines is also attainedby chiral phosphine palladium complexes resulting in enantioselectiveformation of amino alcohols (Hori, et al., J. Org. Chem. 64:5017(1999)). Alternatively, optically pure 1-aryl substituted amino alcoholsare obtained by selective ring opening of corresponding chiral epoxyalcohols with desired amines (Canas et al., Tetrahedron Lett. 32:6931(1991)).

Several methods are known for diastereoselective synthesis of1,3-disubstituted aminoalcohols. For example, treatment of(E)-N-cinnamyltrichloroacetamide with hypochlorous acid results intrans-dihydrooxazine which is readily hydrolysed toerythro-β-chloro-γ-hydroxy-γ-phenylpropanamine in highdiastereoselectivity (Commercon et al., Tetrahedron Lett. 31:3871(1990)). Diastereoselective formation of 1,3-aminoalcohols is alsoachieved by reductive amination of optically pure 3-hydroxy ketones(Haddad et al., Tetrahedron Lett. 38:5981 (1997)). In an alternateapproach, 3-aminoketones are transformed to 1,3-disubstitutedaminoalcohols in high stereoselectivity by a selective hydride reduction(Barluenga et al., J. Org. Chem. 57:1219 (1992)).

Synthesis of Phosphorylation Precursors:

Synthesis of phosphorylation precursors is divided in to two sections:(a) synthesis of P(III) phosphorylation precursors, and (b)stereoselective synthesis of P(V) phosphorylation precursors.

Synthesis of P(I) Phosphorylation Precursors:

Phosphorylation of 5′-alcohol is achieved using cyclic 1′,3′-propanylesters of phosphorylating agents where the agent is in the P(I)oxidation state. One preferred phosphorylating agent is a chlorophospholane (L′=chloro). Cyclic chlorophospholanes are prepared undermild conditions by reaction of phosphorus trichloride with substituted1,3-diols (Wissner, et al, J. Med. Chem. 35:1650 (1992)). Alternativelyphosphoramidites can be used as the phosphorylating agent (Beaucage, etal., Tetrahedron 49:6123 (1993)). Appropriately substitutedphosphorarnidites can be prepared by reacting cyclic chlorophospholaneswith N,N-dialkylamine (Perich, et al., Aust. J. Chem. 43:1623 (1990);Perich, et al, Synthesis 2:142 (1988)) or by reaction of commerciallyavailable dialkylaminophosphorochloridate with substitutedpropane-1,3-diols.

Synthesis of P(V) Phosphorylation Precursors:

In general, synthesis of phosphate esters is achieved by coupling thealcohol with the corresponding activated phosphate precursor forexample, Chlorophosphate (L′=chloro) condensation with 5′-hydroxy ofnucleoside is a well known method for preparation of nucleosidephosphate monoesters. The activated precursor can be prepared by severalwell known methods. Chlorophosphates useful for synthesis of theprodrugs are prepared from the substituted-1,3-propanediol (Wissner, etal, J. Med. Chem. 35:1650 (1992)). Chlorophosphates are made byoxidation of the corresponding chlorophospholanes (Anderson, et al, J.Org. Chem. 49:1304 (1984)), which are obtained by reaction of thesubstituted diol with phosphorus trichloride. Alternatively, thechlorophosphate agent is made by treating substituted-1,3-diols withphosphorus oxychloride (Patois, et al, J. Chem. Soc. Perkin Trans.I:1577 (1990)). Chlorophosphate species may also be generated in situfrom corresponding cyclic phosphites (Silverburg, et al., TetrahedronLett. 37:771 (1996)), which in turn can be either made from achlorophospholane or phosphoramidate intermediate. Phosphorofluoridateintermediate prepared either from pyrophosphate or phosphoric acid mayalso act as precursor in preparation of cyclic prodrugs (Watanabe etal., Tetrahedron Lett. 29:5763 (1988)).

Phosphoramidates (L′=NRR′) are also well-known intermediates for thesynthesis of phosphate esters. Monoalkyl or dialkylphosphoramidate(Watanabe, et al, Chem Pharm Bull 38:562 (1990)),triazolophosphoramidate (Yamakage, et al., Tetrahedron 45:5459 (1989))and pyrrolidinophosphoramidate (Nakayama, et al, J. Am. Chem. Soc.112:6936 (1990)) are some of the known intermediates used for thepreparation of phosphate esters. Another effective phosphorylatingprocedure is a metal catalyzed addition of cyclic chlorophosphate adductof 2-oxazolone. This intermediate attains high selectivity inphosphorylation of primary hydroxy group in presence of secondaryhydroxyl group (Nagamatsu, et al, Tetrahedron Lett. 28:2375 (1987)).These agents are obtained by reaction of a chlorophosphate with theamine or alternatively by formation of the corresponding phosphoramiditefollowed by oxidation.

Synthesis of Enantiomerically Enriched P(V) Phosphorylation Precursors:

The enantioenriched activated phosphorylating agent is synthesized byphosphorylation of an enantioenriched 1-(V)-1,3-propanediol withphosphorodichloridates of formula L-P(O)Cl₂ in the presence of a base(Ferroni, et al., J. Org. Chem. 64:4943 (1999)). Phosphorylation of anenantiomerically pure substituted diol with, for example, a commerciallyavailable phosphorodichloridate R—OP(O)Cl₂, where RO is a leaving group,preferably aryl substituted with electron withdrawing groups, such as anitro or a chloro, produces two diastereomeric intermediates. Therelative configuration of the phosphorus atom is easily determined bycomparison of the ³¹P NMR spectra. The chemical shift of the equatorialphosphoryloxy moiety (trans-isomer) is always more upfield than the oneof the axial isomer (cis-isomer) (Verkade, et al, J. Org. Chem. 42:1549(1977)). These diastereomers can be further equilibrated to give atrans-2,4-substituted phosphorylating agents in presence of a base suchas triethyl amine or DBU. The equilibration to complete inversion of2,4-cis-diastereomer is also achieved in presence of appropriatelysubstituted sodium phenoxide. The equilibration step results in greaterthan 95% ee of the isolated trans-phosphorylating agent.

Synthesis of Nucleosides

All nucleoside moieties of Formula I and Formula II are well describedin the literature. 2′-C-methyl-adenosine and 2′-C-methyl-guanosineanalogs are made by Lewis acid catalyzed reactions of the persilylatedbase and 1′-acetate or benzoate sugar intermediate (Walton et al., J.Am. Chem. Soc. 88:4524 (1966); Harry-O'Kuru et al., J. Org. Chem.62:1754 (1997); WO 01/90121). The 7-deaza analogs are made as describedearlier from 1′-bromo sugar intermediate via reaction of sodium salt ofthe bases (US2002-0147160A1 or WO 02/057827). The glycosylation productsare subjected to deprotection and amination via ammonolysis reaction.

The nucleoside moieties and derivatives thereof of the compounds of thepresent invention may be synthesized by many well-established generalmethods described in the nucleoside literature. Several nucleosidesanalogs described herein are synthesized as illustrated in WO 04/046331and by the methods cited therein. These compounds of the presentinvention can also be made from a wide variety of commercial basesutilizing the 2′-methyl riboglycosylation precursor (US2002-0147160A1 orWO 02/057827) via a range of well-known glycosylation reactions(Vorbruggen and Ruh-Pohlenz, Handbook of Nucleoside Synthesis, Wiley,New York (2001)). Furthermore, deaza and aza nucleoside analogs may beprepared utilizing the methods reported in the case of correspondingribo-analogs by glycosylation with 2′-methyl glycosylation precursor(Robins, et al., Advances in Antiviral Drug Design, Vol. 1, De Clercq,ed., JAI Press, Greenwich, Conn. (1993), pp 39-85). In addition, newbase analogs of the nucleosides can be synthesized by modification ofthe available nucleosides or via synthesis of new bases followed byglycosylation (Chemistry of Nucleosides and Nucleotides, Vols. 1-3,Townsend, ed., Plenum, New York (1988); and Nucleic Acid Chemistry,Vols. 1-4, Townsend and Tipson Eds., Wiley, New York (1986)).

Synthesis of Prodrugs via Coupling of Nucleosides and Prodrug Moiety.

The following procedures on the preparation of prodrugs illustrate thegeneral procedures used to prepare the NMP prodrugs. Prodrug moietiescan be introduced at different stages of the synthesis. Most often theyare made at a later stage, because of the general sensitivity of thesegroups to various reaction conditions. Optically pure prodrugscontaining a single isomer at the phosphorus center are made by couplingof enantiomerically enriched activated phosphate intermediates.

All the procedures described herein, where Y′ and Y″ of —P(O)Y′R¹¹Y″R¹¹of Z″ are oxygen, are also applicable for the preparation of theprodrugs where they are NRV by appropriate substitution or protection ofnitrogen.

The preparation of prodrugs is further organized into (1) synthesis viaP(III) intermediates, (2) synthesis via P(V) intermediates, and (3)miscellaneous methods.

Synthesis of Prodrugs via P(III) Intermediates:

wherein Q is N or CH; and L is H and M is NH₂ or M is OH and L is NH₂.

Chlorophospholanes are used to phosphorylate alcohols on nucleosides inthe presence of an organic base (e.g., triethylamine, pyridine).Alternatively, the phosphite can be obtained by coupling the nucleosidewith a phosphoramidate in the presence of a coupling promoter such astetrazole or benzimidazolium triflate (Hayakawa et al., J. Org. Chem.,1996, 61, 7996). Phosphite diastereomers may be isolated by columnchromatography or crystallization (Wang, et al, Tetrahedron Lett 38:3797(1997); Bentridge et al., J. Am. Chem. Soc. 111:3981 (1989)). Sincecondensation of alcohols with chlorophospholanes or phosphoramidites isan S_(N)2(P) reaction, the product is expected to have an invertedconfiguration. This allows for the stereoselective synthesis of cyclicphosphites. Isomeric mixtures of phosphorylation reactions can also beequilibrated (e.g., thermal equilibration) to a more thermodynamicallystable isomer.

The resulting phosphites are subsequently oxidized to the correspondingphosphate prodrugs using an oxidant such as molecular oxygen ort-butylhydroperoxide (Meier et al., Bioorg, Med. Chem. Lett. 7:1577(1997)). Oxidation of optically pure phosphites is expected tostereoselectively provide optically active prodrugs (Mikolajczyk, etal., J. Org. Chem. 43:2132 (1978). Cullis, P. M., J. Chem. Soc., ChemCommun. 1510 (1984); Verfurth, et al., Chem. Ber. 129:1627 (1991)).

Synthesis of Prodrugs via P(V) Intermediates:

For the synthesis of cis-prodrugs of Formula I and Formula II, theprodrug moiety can be introduced at different stages of the synthesis.Most often the cyclic phosphates are introduced at a later stage,because of the general sensitivity of these groups to various reactionconditions. The synthesis can also proceed through using a protected orunprotected nucleoside or nucleoside analog depending on the reactivityof the functional groups present in the compound. Single stereoisomersof the cis-prodrugs can be made either by separation of thediastereoisomers/enantiomers by a combination of column chromatographyand/or crystallization, or by enantiospecific or enantioselectivesynthesis using enantioenriched activated phosphate intermediates.

Synthesis of Enantiomerically Enriched Prodrugs:

wherein Q is N or CH; and L is H and M is NH₂ or M is OH and L is NH₂.

The general procedure for the phosphorylation of protected nucleosidesis accomplished by reacting a suitably protected nucleoside with a baseand reacting the alkoxide generated with the phosphorylating reagent.The protected nucleoside can be prepared by one skilled in the art usingone of the many procedures described for the protection of nucleosides(Greene T. W., Protective Groups in Organic Chemistry, John Wiley &Sons, New York (1999)). The nucleoside is protected in such a way as toexpose the hydroxyl group on which to add the phosphate group whileprotecting all the remaining hydroxyls and other functional groups onthe nucleoside that may interfere with the phosphorylation step or leadto regioisomers. In one aspect, the protecting groups selected areresistant to strong bases, e.g., ethers, silyl ethers and ketals. In oneaspect, the protecting groups are optionally substituted MOM ethers, MEMethers, trialkylsilyl ethers and symmetrical ketals. In another aspect,the protecting groups are t-butyldimethylsilyl ether and isopropylidene.Further protection entails masking of the amino group of the basemoiety, if present, so as to eliminate any acidic protons. In one aspectthe selected N-protecting groups are selected from the groups of dialkylformamidines, mono and dialkyl imines, mono and diaryl imines. In oneaspect, the N-protecting groups are selected from the groups of dialkylformamidines and mono-alkyl imine and mono aryl imine. In one aspect themono-alkyl imine is benzylimine and the mono-aryl imine is phenylimine.In another aspect, the N-protecting group is a symmetrical dialkylformamidine selected from the group of dimethyl formamidine and diethylformamidine.

Generation of the alkoxide of the exposed hydroxyl group on the suitablyprotected nucleoside is accomplished with a base in an aprotic solventthat is not base sensitive such as THF, dialkyl and cyclic formamides,ether, toluene and mixtures of those solvents. In one aspect, thesolvents are DMF, DMA, DEF, N-methylpyrrolidinone, THF, and mixture ofthose solvents.

Many different bases have been used for the phosphorylation ofnucleosides and non-nucleoside compounds with cyclic and acyclicphosphorylating agents. For example trialkylamines such as triethylamine(Roodsari et al., J. Org. Chem. 64:7727 (1999)) orN,N-diisopropylethylamine (Meek et al., J. Am. Chem. Soc. 110:2317(1988)); nitrogen containing heterocyclic amines such as pyridine(Hoefler et al., Tetrahedron 56(11), 1485 (2000)), N-methylimidazole(Vankayalapati et al., J. Chem. Soc. Perk T 1 14:2187 (2000)),1,2,4-triazole (Talcaku et al., Chem. Lett. (5):699 (1986)) or imidazole(Dyatkina et al., Tetrahedron Lett. 35:1961 (1994)); organometallicbases such as potassium t-butoxide (Postel et al., J. Carbohyd. Chem.19:171 (2000)), butyllithium (Torneiro et al., J. Org. Chem. 62:6344(1977)), t-butylmagnesium chloride (Hayakawa et al., Tetrahedron Lett.28:2259 (1987)) or LDA (Aleksiuk et al., J. Chem. Soc. Chem. Comm. (1),11 (1993)); inorganic bases such as cesium fluoride (Takaku, et al.,Nippon Kagaku Kaishi (10), 1968 (1985)), sodium hydride (Hanaoka et al.,Heterocycles 23:2927 (1985)), sodium iodide (Stromberg, et al., J.Nucleos. Nucleot. 6:815 (1987)), iodine (Stromberg, et al., J. Nucleos.Nucleot. 6:815 (1987)) or sodium hydroxide (Attanasi, et al., PhosphorusSulfur 35:63 (1988)); metals such as copper (Bhatia, et al., TetrahedronLett. 28:271 (1987)). However, no reaction or racemization at thephosphorus stereogenic center was observed when coupling ofphosphorylating reagent was attempted using the previously describedprocedures. Especially, no reaction was observed with bases previouslyused with substituted cyclic phosphorylating agent to give thecorresponding cyclic phosphate in high yield such as sodium hydride(Thuong et al., Bull. Soc. Chim. Fr. 667 (1974)), pyridine(Ayral-Kaloustian et al., Carbohydr. Res. 187 (1991)), butyl-lithium(Hulst et al., Tetrahedron Lett. 1339 (1993)), DBU (Merckling et al.,Tetrahedron Lett. 2217 (1996)), triethylamine (Hadvary et al., Helv.Chim. Acta 69:1862 (1986)), N-methylimidazole (Li, et al., TetrahedronLett. 6615 (2001)) or sodium methoxide (Gorenstein et al., J. Am. Chem.Soc. 5077 (1980)). It was found that the use of Grignard reagentspromoted phosphorylation with minimal epimerization of the phosphoruscenter. In one aspect, Grignard reagents are alkyl and aryl Grignards.In another aspect, the Grignard reagents are t-butyl magnesium halidesand phenyl magnesium halides. In another aspect, the Grignard reagentsare t-butylmagnesium chloride and phenylmagnesium chloride.

In another aspect magnesium alkoxides are used to generate the magnesium5′-alkoxide of the nucleoside. In one aspect magnesium alkoxides areselected from the group of Mg(O-t-Bu)₂, and Mg(O-iPr)₂.

The protected prodrugs generated as described above are then subjectedto a deprotection step to remove all the protecting groups using one ofthe many methods known to those skilled in the art (Greene, T. W.,Protective Groups in Organic Chemistry, John Wiley & Sons, New York(1999)) and that are compatible with the stability of the phosphateprodrug. In one aspect, deprotection reagents include fluoride salts toremove silyl protecting groups, mineral or organic acids to remove acidlabile protecting groups such as silyl and/or ketals and N-protectinggroups, if present. In another aspect, reagents are tetrabutylammoniumfluoride (TBAF), hydrochloric acid solutions and aqueous TFA solutions.Isolation and purification of the final prodrugs, as well as allintermediates, are accomplished by a combination of columnchromatography and/or crystallization.

The sequence provides methods to synthesize single isomers of compoundsof Formula I and Formula II. For compounds of Formula II, due to thepresence of a stereogenic center at the carbon where V is attached onthe cyclic phosphate reagent, this carbon atom can have two distinctorientations, namely R or S. As such the trans-phosphate reagentprepared from a racemic diol can exist as either the S-trans or R-transconfiguration and results in a S-cis and R-cis prodrug mixture. Thereaction of the C′-S-trans-phosphate reagent generates theC′-S-cis-prodrug of the nucleoside while reaction with theC′-R-trans-phosphate reagent generates the C′-R-cis-prodrug.

Miscellaneous Phosphorylation Methods:

Coupling of activated phosphates with alcohols is accomplished in thepresence of an organic base. For example, chlorophosphates synthesizedas described in the earlier section react with an alcohol in thepresence of a base such as pyridine or N-methylimidazole. In some casesphosphorylation is enhanced by in situ generation of iodophosphate fromchlorophosphate (Stomberg, et al., Nucleosides & Nucleotides. 5:815(1987)). Phosphorofluoridate intermediates have also been used inphosphorylation reactions in the presence of a base such as CsF orn-BuLi to generate cyclic prodrugs (Watanabe et al., Tetrahedron Lett.29:5763 (1988)).

Phosphoramidate intermediates are known to couple by transition metalcatalysis (Nagamatsu, et al., Tetrahedron Lett. 28:2375 (1987)).

Reaction of the optically pure diastereomer of phosphoramidateintermediate with the hydroxyl of nucleoside in the presence of an acidproduces the optically pure phosphate prodrug by direct S_(N)2(P)reaction (Nakayama, et al., J. Am. Chem. Soc. 112:6936 (1990)).Alternatively, reaction of the optically pure phosphate precursor with afluoride source, preferably cesium fluoride or TBAF, produces the morereactive phosphorofluoridate which reacts with the hydroxyl of thenucleoside to give the optically pure prodrug by overall retention ofconfiguration at the phosphorus atom (Ogilvie, et al., J. Am. Chem.Soc., 99:1277 (1977)).

Prodrugs of Formula I are synthesized by reaction of the correspondingphosphodichloridate and an alcohol (Khamnei, et al., J. Med. Chem.39:4109 (1996)). For example, the reaction of a phosphodichloridate withsubstituted 1,3-diols in the presence of base (such as pyridine andtriethylamine) yields compounds of Formula I.

Such reactive dichloridate intermediates can be prepared from thecorresponding acids and the chlorinating agents such as thionyl chloride(Starrett, et al, J. Med. Chem. 1857 (1994)), oxalyl chloride (Stowell,et al., Tetrahedron Lett. 31:3261 (1990)), and phosphorus pentachloride(Quast, et al., Synthesis 490 (1974)).

Phosphorylation of an alcohol is also achieved under Mitsunobu reactionconditions using the cyclic 1′,3′-propanyl ester of phosphoric acid inthe presence of triphenylphosphine and diethyl azodicarboxylate (Kimura,et al., Bull. Chem. Soc. Jpn. 52:1191 (1979)). The procedure can beextended to prepare enantiomerically pure phosphates from thecorresponding phosphoric acids. Phosphate prodrugs are also preparedfrom the free acid by Mitsunobu reactions (Mitsunobu, Synthesis, 1(1981); Campbell, J. Org. Chem. 52:6331 (1992)), and other acid couplingreagents including, but not limited to, carbodiimides (Alexander, etal., Collect. Czech. Chem. Commun. 59:1853 (1994); Casara, et al.,Bioorg. Med. Chem. Lett. 2:145 (1992); Ohashi, et al., Tetrahedron Lett.29:1189 (1988)), and benzotriazolyloxytris-(dimethylamino)phosphoniumsalts (Campagne, et al., Tetrahedron Lett. 34:6743 (1993)).Cyclic-1,3-propanyl prodrugs of phosphates are also synthesized from NMPand substituted propane-1,3-diols using a coupling reagent such as1,3-dicyclohexylcarbodiimide (DCC) in presence of a base (e.g.,pyridine). Other carbodiimide based coupling agents such as1,3-diisopropylcarbodiimide and the water soluble reagent,1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI) canalso be utilized for the synthesis of cyclic prodrugs.

Phosphate prodrugs can be prepared by an alkylation reaction between thephosphate corresponding tetrabutylammonium salts andsubstituted-1,3-diiodopropanes made from 1,3-diols (Farquhar, et al.,Tetrahedron Lett. 36:655 (1995)). Furthermore, phosphate prodrugs can bemade by conversion of nucleoside to the dichloridate intermediate withphosphoryl chloride in presence of triethylphosphite and quenching withsubstituted-1,3-propanediols (Farquharet al., J. Org. Chem. 26:1153(1983)).

Phosphorylation can also be achieved by making the mixed anhydride ofthe cyclic diester of phosphoric acid and a sulfonyl chloride,preferably 8-quinolinesulfonyl chloride, and reacting the hydroxyl ofthe nucleoside in the presence of a base, preferably N-methylimidazole(Takaku, et al, J. Org. Chem. 47:4937 (1982)). In addition, startingfrom an enantiomerically pure cyclic diester of a phosphoric acid,obtained by resolution (Wynberg, et al., J. Org. Chem. 50:4508 (1985)),one can obtain enantiomerically pure phosphates.

Synthesis of compounds of Formula I, wherein Z″ is P(O)Y′R¹¹Y″R¹¹ andeach R¹¹ is independently substituted by acyclic groups, Y′ and Y″ areeach independently selected from the group consisting of —O—, and—NR^(v)— may be also attained by the procedures described above for thecyclic prodrugs.

Synthesis of 6-substituted Prodrugs:

Prodrugs at the 6-position may be prepared from the corresponding haloderivatives of the nucleosides. The prodrug substitution is made at thenucleoside stage (before 5′-prodrug formation) from the correspondingchloro or hydroxy functionalities in the case of compounds of in which Bis a purine base or purine base derivative substituted at the 6-position(e.g., N₃, H, —COR, CO₂R). Synthesis of such nucleoside precursors areattained as described earlier (WO 02/057287). Preparation of thesepurine analogs by azido displacement (Aso et al., J. Chem Soc PerkinTrains II 8:1637 (2000)) or hydrogention (Freer et al., Tetrahedron56:45 (2000)) are well known methods. Subsequently, these prodrugfunctionality substituted nucleosides are transformed to correspondingmonophosphate cyclic prodrugs of Formula I or Formula II.

EXAMPLES

The compounds used in this invention and their preparation can beunderstood further by the Examples, which illustrate some of theprocesses by which these compounds are prepared. These Examples shouldnot however be construed as specifically limiting the invention, andvariations of the compounds, now known or later developed, areconsidered to fall within the scope of the present invention ashereinafter claimed.

Compounds of Formula I are prepared as outlined below. The TLCconditions given are utilizing plates of Analtech UNIPLATE, silica gelGHLF, scored 10×20 cm, 250 micron.

Synthesis of Racemic 1-(aryl)propane-1,3-diols:

Example 1 Preparation of 1-(2′-Furanyl)-Propane-1,3-Diol via GrignardAddition and Hydroboration

To a solution of 2-furaldehyde (3 g, 31.2 mmol) in THF (60 mL) was added1 M vinyl magnesium bromide in THF (34 mL) at 0° C. After stirring foran hour, a solution of 1 M BH₃.THF complex in THF was added. Thereaction was quenched with 3N NaOH (20 mL) and 30% hydrogen peroxide (10mL) at 0° C. The organic fraction was separated and concentrated. Thecrude product was chromatographed by eluting with 5%methanol-dichloromethane to give 1-(2′-furyl)propane-1,3-diol (1 g).

Example 2 Preparation of 1-(2′-Pyridyl)-Propane-1,3-Diol via BenzylicOxidation

Step A: (J. Org. Chem. 22:589 (1957))

To a solution of 3-(2′-pyridyl)propan-1-ol (10 g, 72.9 mmol) in aceticacid (75 mL) was added 30% hydrogen peroxide slowly. The reactionmixture was heated to 80° C. for 16 h. The reaction was concentratedunder vacuum and the residue was dissolved in acetic anhydride (100 mL)and heated at 110° C. overnight. Acetic anhydride was evaporated uponcompletion of the reaction. Chromatography of the mixture by elutingwith methanol-methylene:chloride (1:9) resulted in 10.5 g of purediacetate.

Step B:

To a solution of diacetate (5 g, 21.1 mmol) in methanol-water (3:1, 40mL) was added potassium carbonate (14.6 g, 105.5 mmol). After stirringfor 3 h at room temperature, the reaction mixture was concentrated. Theresidue was chromatographed by eluting with methanol-methylene chloride(1:9) to give 2.2 g of crystalline diol.

Example 3 Preparation of 1-(Aryl)-Propane-1,3-Diol from Propane-1,3-Diolvia Grignard Addition

Step A: (J. Org. Chem. 53:911 (1988))

To a solution of oxalyl chloride (5.7 mL, 97 mmol) in dichloromethane(200 mL) at −78° C. was added dimethyl sulfoxide (9.2 mL, 130 mmol). Thereaction mixture was stirred at −78° C. for 20 min before addition of3-(benzyloxy)propan-1-ol (11 g, 65 mmol) in dichloromethane (25 mL).After an hour at −78° C., reaction was quenched with triethylamine (19mL, 260 mmol) and warmed to room temperature. Work-up and columnchromatography by elution with dichloromethane resulted in 8 g of3-(benzyloxy)propan-1-al.

Step B:

To a solution of 3-(benzyloxy)propan-1-al (1 g, 6.1 mmol) in THF at 0°C. was added a 1 M solution of 4-fluorophenylmagnesium bromide in THF(6.7 mL, 6.7 mmol). The reaction was warmed to room temperature andstirred for 1 h. Work-up and column chromatography by elution withdichloromethane resulted in 0.7 g of alcohol.

Step C:

To a solution of benzyl ether (500 mg) in ethyl acetate (10 mL) wasadded 10% Pd(OH)₂C (100 mg). The reaction was stirred under hydrogen gasfor 16 h. The reaction mixture was filtered through Celite andconcentrated. Chromatography of the residue by elution with ethylacetate-dichloromethane (1:1) resulted in 340 mg of product.

Example 4 General Procedure for Preparation of 1-ArylSubstituted-Propane-1,3-Diol from Aryl Aldehyde

Step A: (J. Org. Chem. 55:4744 (1990))

To a −78° C. solution of diisopropylamine (2 mmol) in THF (0.7 mm/mmoldiisopropylamine) was slowly added n-butyllithium (2 mmol, 2.5 Msolution in hexanes). The reaction was then stirred for 15 min at −78°C. before a solution of ethyl acetate (2 mmol) in THF (0.14 mmol ethylacetate) was slowly introduced. After stirring an additional 30 min at−78° C., a THF solution containing the aryl aldehyde (1.0 mmol in 0.28mL THF) was added. The reaction was then stirred at −78° C. for 30 min,warmed to room temperature and stirred an additional 2 h. After aqueouswork up (0.5 M HCl), the organic layer was concentrated to a crude oil(beta-hydroxyester).

Step B:

The crude hydroxyester was dissolved in ether (2.8 mL/mmol), cooled toice bath temperature, and lithium aluminum hydride (3 mmol) was addedbatch wise. The reaction was stirred allowing the cooling bath to meltand the reaction to reach room temperature. After stirring overnight atroom temperature, the reaction was cooled back to ice bath temperatureand quenched with ethyl acetate. Aqueous work up (0.5 M HCl) affordedthe crude diol, which was purified either by chromatography ordistillation.

Example 4a Synthesis of 1-(3-methoxycarbonylphenyl)-1,3-propanediol

1-(3-bromophenyl)-1,3-propane diol was prepared as Example 4 and furtherderivatized as follows:

A pressure vessel was charged with 1-(3-bromophenyl)-1,3-propanediol (2g, 8.6 mmol), methanol (30 mL), triethylamine (5 mL) andbis(triphenylphosphine)palladium dichloride (0.36 g, 05 mmol). Thesealed vessel was pressurize with carbon monoxide at 55 psi and heatedat 85° C. for 24 h. The cooled vessel was opened and the reactionmixture was filtered through Celite and rinsed with methanol. Thecombined filtrates were concentrated under reduced pressure and theresidue was purified by column chromatography (silica gel, hexanes/ethylacetate 1/1) to afford the title compound (1.2 g)

TLC: hexanes/ethyl acetate 2/8; Rf=0.5

¹H NMR (CDCl₃, Varian Gemini 200 MHz): 5.05-4.95 (m, 1H), 3.9 (s, 3H),2-1.8 (m, 2H).

Example 4b Synthesis of 1-(4-methoxycarbonylphenyl)-1,3-propanediol

1-(4-bromophenyl)-1,3-propane diol was prepared as Example 4 and furtherderivatized as Example 4a.

TLC: hexanes/ethyl acetate 3/7; Rf=0.35

¹H NMR (CDCl₃, Varian Gemini 200 MHz): 5.1-5 (m, 1H), 3.91 (s, 3H),2.05-1.9 (m, 2H).

Synthesis of Enantioenriched 1-(aryl)-propane-1,3-diols:

Example 5 General Procedure for Resolution of Racemic 1,3-diols

Racemic diols synthesized as in Examples 1-4 may be resolved to yieldboth enantiomers as described in the following procedure.

Step A:

To a solution of diol (1.0 mmol) in THF (1.0 ml) was addedhexamethyldisilazane (2.1 mmol) followed by a catalytic amount oftrimethylsilyltriflate (2-3 drops). After stirring at room temperaturefor 1 h, the reaction was diluted with hexane (4 mL) and subjected towork up with ice-cold water. The resulting disilylether was eitherpurified by chromatography or, if sufficiently pure, used crude in thenext reaction.

Step B:

To a solution of disilylether (1.0 mmol) and (−)-menthone (1.1 mmol) indichloromethane (2.0 ml) at −40° C., was slowly addedtrimethylsilyltriflate (0.11 mmol). The reaction was then kept at −50°to −60° C. for 48 h, at which time pyridine was added to quench thereaction. After warming to room temperature, the crude mixture wasdiluted with hexane (4.0 ml) and subjected to aqueous work up. The twoketals were separated by chromatography.

Step C:

The separated ketals were hydrolyzed by adding a catalytic amount ofconcentrated hydrochloric acid to a methanol (4.0 mL/mmol) solution ofeach. After stirring overnight at room temperature, the methanol wasremoved under vacuum and the residue was subjected to aqueous work up.The resolved diols were further purified by either chromatography ordistillation.

Example 6 Synthesis of Enantioenriched1-(3′-chlorophenyl)-1,3-dihydroxypropane via Sharpless AsymmetricEpoxidation Step A:

To a dispersion of m-chloro-cinnamic acid (25 g, 137 mmol) in ethanol(275 mL) was added conc. sulfuric acid (8 mL) at room temperature. Thereaction was refluxed overnight and concentrated. Ice-cold water wasadded to the crude and precipitated white solid was filtered and washedwith cold water. The precipitate was dried under vacuum overnight togive 25 g of ester. (Rf=0.5 in dichloromethane on silica)

Step B:

To a solution of ethyl-m-chlorocinnamate (23 g, 109.5 mmol) indichloromethane at −78° C. was added 1 M DIBAL-H in dichloromethane (229mL, 229 mmol) dropwise over 1 h. The reaction was stirred at −78° C. foran additional 3 h. Ethylacetate was added to quench excess DIBAL-H andsaturated aq. potassium sodium tartrate was added and the reaction wasstirred at room temperature for 3 h. The organic layer was separated andsalts were washed with ethyl acetate. The combined organic extracts wereconcentrated and distilled at 120° C./0.1 mm to give 14 g of pureallylic alcohol. (Rf=0.38 in 1:1 ethylacetate:hexane on silica)

Step C:

To a solution of m-chlorocinnamyl alcohol (5 g, 29.76 mmol) indichloromethane (220 mL) was added activated 4 Å molecular sieves powder(2.5 g) and the mixture was cooled to −20° C. (+)-Diethyl tartrate (0.61mL, 3.57 mmol) was added at −20° C. and stirred for 15 min before addingtitanium tetraisopropoxide (0.87 g, 2.97 mmol). The reaction was stirredfor additional 30 min and 5-6 M solution of t-butylhydroperoxide inheptane (10 mL, 60 mmol) was added dropwise while maintaining theinternal temperature at −20 to −25° C. The mixture was stirred for anadditional 3 h at −20° C. and a 10% sodium hydroxide in saturated aq.sodium chloride (7.5 mL) followed by ether (25 mL) were added. Thereaction was warmed to 10° C. and stirred for 15 min before addinganhydrous magnesium sulfate (10 g) and Celite (1.5 g). The mixture wasfurther stirred for additional 15 min, filtered and concentrated at 25°C. to give crude epoxy alcohol. (Rf=0.40 in 1:1 ethylacetate:hexane onsilica).

Step D:

To a solution of crude m-chloroepoxycinnamyl alcohol obtained fromearlier reaction in dimethoxyethane (300 mL) was added a 65% Red-Alsolution in toluene (18.63 mL, 60 mmol) dropwise under nitrogen at 0° C.After stirring at room temperature for 3 h, the solution was dilutedwith ethyl acetate (400 mL) and quenched with aq. saturated sodiumsulfate solution (50 mL). After stirring at room temperature for 30 min,the resulting white precipitate formed was filtered and washed withethylacetate. The filtrate was dried and concentrated. The crude productwas distilled at 125-130° C./0.1 mm to give 3.75 g of enantioenriched(R)-1-(3′-chlorophenyl)-1,3-dihydroxypropane. (Rf=0.40 in 1:1ethylacetate:dichloromethane).

Enantiomeric excesses were defined as diacetates (prepared by treatmentof diols with acetic anhydride, triethylamine, cat.DMAP indichloromethane) by HPLC ((S,S) Whelko-0, 250 cm×4.0 mm ID purchasedfrom Regis).

(R)-1-(3′-chlorophenyl)-1,3-dihydroxypropane: 91% ee

(+)Diisopropyltartrate provided >96% ee in(R)-1-(3′-chlorophenyl)-1,3-dihydroxypropane.

(S)-1-(3′-chlorophenyl)-1,3-dihydroxypropane was also prepared underidentical conditions via asymmetric epoxidation and reduction protocolutilizing (−)-tartrate in similar yields.(S)-3-(3′-chlorophenyl)-1,3-dihydroxypropane was obtained with 79% ee.

Example 7 Synthesis of Enantioenriched1-(3′-chlorophenyl)-1,3-dihydroxypropane via Hydrogen Transfer Reaction

Step A: Preparation of methyl 3-(3′-chlorophenyl)-3-oxo-propanoate:

A 22 L, 3-neck round bottom flask was equipped with a mechanicalstirrer, thermowell/thermometer and nitrogen inlet (bubbler in-line).The flask was flushed with nitrogen and charged sequentially with THF (6L), potassium t-butoxide (1451 g), and THF (0.5 L). The resultingmixture was stirred at ambient temperature for 15 min. and a 20° C.water bath was applied. A 3 L round bottom flask was charged with3′-chloroacetophenone (1000 g) and diethylcarbonate (1165 g), and theresulting yellow solution was added slowly to the stirred potassiumt-butoxide solution, maintaining the temperature between 16 and 31° C.After the addition was complete (1 h, 10 min.), the cooling bath wasremoved and the solution was stirred for 1 h, 30 min. TLC indicated thatthe reaction was complete. A 5 gallon stationary separatory funnel wascharged with ice water (4 L) and concentrated hydrochloric acid (1.3 Lof 12 M solution). The dark red reaction solution was quenched into theaqueous acid and the mixture was stirred for 15 min. The layers wereseparated and the aqueous phase (lower) was extracted again with toluene(4 L). The combined organic extracts were washed with saturated brine(2×3 L, 10 min. stirring time each), dried MgSO₄), filtered andconcentrated under reduced pressure to provide 1480 g of a brown oil.The oil was placed under high vacuum (10 torr) overnight to give 1427 g.The material was vacuum distilled (short path column, fraction cutterreceiver) and the fraction at 108-128° C./1-0.5 torr was collected toprovide 1273.9 g of a yellow oil. (Rf=0.36 in 20% ethylacetate/hexanes).

Step B: Preparation of methyl(S)-3-(3′-chlorophenyl)-3-hydroxypropionate:

A 12 L, 3-neck round bottom flask was equipped with a mechanicalstirrer, thermometer, addition funnel (500 mL) and nitrogen inlet(bubbler in-line). The flask was flushed with nitrogen and charged withformic acid (292 mL, 350 g). Triethylamine (422 mL, 306 g) was chargedto the addition funnel, then added slowly with stirring, maintaining thetemperature less than 45° C. After the addition was complete (1 h, 30min), the solution was stirred with the ice bath applied for 20 min.,then at ambient temperature for an additional 1 h. The flask was chargedsequentially with methyl 3-(3-chlorophenyl)-3-oxo-propanoate (1260 g),DMF (2.77 L including rinsing volume) and (S,S)-Ts-DPEN-Ru—Cl-(p-cymene)(3.77 g). The flask was equipped with a heating mantle and the additionfunnel was replaced with a condenser (5 C circulating coolant forcondenser). The stirred reaction solution was slowly heated to 60° C.(90 min. to attain 60° C.) and the contents were maintained at 60° C.for 4.25 h. HPLC indicated 3% starting material remained. The solutionwas stirred at 60° C. for an additional 8 h, then gradually cooled toambient temperature overnight. HPLC indicated 0.5% starting material. A5 gallon stationary separatory funnel was charged with water (10 L) andMTBE (1 L). The reaction solution was poured into the aqueous mixtureand the reaction flask was rinsed into the separatory funnel with anadditional 1 L of MTBE. The contents were stirred for several minutesand the layers were separated. The aqueous phase was extracted withadditional MTBE (2×1 L), and the combined organic extracts were washedwith brine (1 L), and concentrated under reduced pressure to provide1334 g of a red oil. The oil was used without further purification forthe next step.

The crude hydroxyester (10 mg, 0.046 mmol) was dissolved indichloromethane (1 mL). Acetic anhydride (22 μL, 0.23 mmol) and4-(dimethylamino)pyridine (22 mg, 0.18 mmol) were added and the solutionwas stirred at ambient temperature for 15 min. The solution was dilutedwith dichloromethane (10 mL) and washed with 1 M hydrochloric acid (3×3mL). The organic phase was dried (MgSO₄), filtered and concentratedunder reduced pressure. The residual oil was dissolved in methanol andanalyzed by chiral IIPLC (Zorbax Rx-C18, 250×4.6 mm; mobile phase: 65/35(v/v) water/acetonitrile, isocratic; flow rate=1.5 mL/min; inj.volume=15 μL; UV detection at 220 nm. Retention times: Product=9.3 min,starting material=17.2 min). The hydroxyester was derivatized to theacetate for analysis by chiral HPLC and shown to give 91% ee. (HPLCconditions: Column: Pirkle covalent (S,S) Whelk-O 10/100 krom FEC,250×4.6 mm; mobile phase: 70/30 (v/v) methanol/water, isocratic; flowrate: 1.5 mL/min; inj. volume=10 μL; UV detection at 220 nm. Retentiontimes: S-hydroxyester (acetate)=9.6 min, R-hydroxyester (acetate)=7.3min.)

Step C: Preparation of (S)-3-(3′-chlorophenyl)-3-hydroxypropanoic Acid:

To the crude hydroxyester in a 10 L rotary evaporator flask was addedsodium hydroxide solution (2.5 L of 2 M solution). The resultingsolution was stirred on the rotary evaporator at ambient pressure andtemperature for 2 h. HPLC indicated 5% starting material still remained(HPLC conditions: Column: Zorbax Rx-C18, 250×4.6 mm; mobile phase: 65/35(v/v) water/acetonitrile, isocratic; flow rate=1.5 mL/min; inj. volume15 μL; UV detection at 220 nm. Retention times: Product=3.8 min,starting material=18.9 min.). The pH of the solution was 11 (wide rangepH paper). Additional 2 M NaOH solution was added to adjust the pH to 14(approx. 100 mL), and the solution was stirred for an additional 30 min.HPLC indicated the reaction was complete. The solution was transferredto a 5 gallon stationary separatory funnel and extracted with MTBE (2L). The layers were separated and the organic extract was discarded. Theaqueous phase was transferred back to the separatory funnel andacidified with 12 M HCl solution (600 mL). The mixture was extractedwith MTBE (1×2 L, 2×1 L). The combined acidic organic extracts weredried (MgSO₄), filtered and concentrated under reduced pressure to give1262 g of a brown, oily semi-solid. The residue was slurried with ethylacetate (1 L) and transferred to a 12 L, 3-neck round bottom flaskequipped with a mechanical stirrer, heating mantle, condenser andthermometer. The stirred mixture was heated to dissolve all solids (28°C.) and the dark solution was cooled to 10° C. (a precipitate formed at11° C.). The mixture was slowly diluted with hexanes (4 L over 1 h) andthe resulting mixture was stirred at <10° C. for 2 h. The mixture wasfiltered and the collected solid was washed with cold 4/1 hexanes/ethylacetate (1 L), and dried to constant weight (−30 in. Hg, 50° C., 4 h).Recovery=837 g of a beige solid. mp=94.5-95.5° C. A 50 mg sample ofhydroxyacid was reduced to the diol with borane-THF (see Step D). Theresulting crude diol was diacetylated (as described in Step B)) andanalyzed by chiral HPLC. Retention times: S-diol (diacetate)=12.4 min,R-diol (diacetate)=8.8 min.) ee=98%

A second crop of hydroxyacid was isolated. The filtrate from above wasconcentrated under reduced pressure to give 260 g of a brown sludge. Thematerial was dissolved in ethyl acetate (250 mL) and the stirred darksolution was slowly diluted with hexanes (1000 mL) and the resultingmixture was stirred at ambient temperature overnight. The mixture wasfiltered and the collected solid was washed with 5/1 hexanes/ethylacetate (200 mL), and dried to constant weight (−30 in. Hg, 50° C., 16h). Recovery=134 g of a beige solid. ee=97%

Step D: Preparation of (s)-(−)-1-(3-chlorophenyl)-1,3-propanediol:

A 22 L, 3-neck round bottom flask was equipped with a mechanicalstirrer, thermowell/thermometer and nitrogen inlet (outlet to bubbler).The flask was charged with 2 M borane-THF (3697 g, 4.2 L) and thestirred solution was cooled to 5° C. A solution of(S)-3-(3-chlorophenyl)-3-hydroxypropanoic acid (830 g) in THF (1245 mL)was prepared with stirring (slightly endothermic). The reaction flaskwas equipped with an addition funnel (1 L) and the hydroxyacid solutionwas slowly added to the stirred borane solution, maintaining thetemperature <16° C. After the addition was complete (3 h), the mixturewas stirred at ice bath temperature for 1.5 h. The reaction was quenchedby careful addition of water (2.5 L). After the addition was complete(30 min), 3 M NaOH solution (3.3 L) was added (temperature increased to35° C.) and the resulting mixture was stirred for an additional 20 min.(temperature=30° C.). The reaction mixture was transferred to a 5 gallonstationary separatory funnel and the layers were separated. The aqueousphase was extracted with MTBE (2.5 L) and the combined organic extracts(THF and MTBE) were washed with 20 wt % NaCl solution (2 L) and stirredwith MgSO₄ (830 g) for 30 min. The mixture was filtered through Celiteand concentrated under reduced pressure to provide 735 g of a thick,brown oil.

The oil was purified by vacuum distillation and the fraction at 135-140°C./0.2 mm Hg was collected to provide 712.2 g of a colorless oil.

The diol was diacetylated and analyzed by chiral HPLC (e.e.=98%) (seeStep B). Retention times: S-diol (diacetate)=12.4 min, R-diol(diacetate)=8.9 min. [α]_(D)=−51.374 (5 mg/mL in CHCl₃)

Example 8 Synthesis of Enantioenriched1-(4′-pyridyl)-1,3-Dihydroxypropane via Hydrogen Transfer Reaction

Step A: Synthesis of methyl 3-oxo-3-(pyridin-4-yl)-propanoate

A 50 L, 3-neck flask was equipped with an overhead stirrer, heatingmantle, and nitrogen inlet. The flask was charged with THF (8 L),potassium t-butoxide (5 kg, 44.6 mol), and THF (18 L). 4-Acetylpyridine(2.5 kg, 20.6 mol) was added, followed by dimethylcarbonate (3.75 L,44.5 mol). The reaction mixture was stirred without heating for 2.5 hthen with heating to 57-60° C. for 3 h. The heat was turned off and themixture cooled slowly overnight (15 h). The mixture was filtered througha 45 cm Buchner funnel. The solid was returned to the 50 L flask anddiluted with aqueous acetic acid (3 L acetic acid in 15 L of water). Themixture was extracted with MTBE (1×16 L, 1×12 L). The combined organiclayers were washed with aqueous Na₂CO₃ (1750 g in 12.5 L water),saturated aqueous NaHCO₃ (8 L), and brine (8 L) then dried over MgSO₄(500 g) overnight (15 h). The solution was filtered and the solventremoved by rotary evaporation to a mass of 6.4 kg. The resultingsuspension was cooled in an ice bath with stirring for 2 h. The solidwas collected by filtration, washed with MTBE (500 mL), and dried in avacuum oven at 20° C. for 15 h, giving 2425 g of the keto ester as apale yellow solid.

The MTBE mother liquor was concentrated to approximately 1 L. Theresulting suspension was cooled in an ice bath for 1 h. The solid wascollected by filtration, washed with MTBE (2×150 mL), and dried in avacuum oven to give 240 g of a second crop.

TLC. Merck silica gel plates, 1:2 THF/hexane, UV lamp, Rf of SM=0.25, Rfof product=0.3.

Melting Point: 74-76° C.

Step B: Synthesis of S-methyl-3-hydroxy-3-(pyridin-4-yl)-propanoate

A 22 L, 3-neck round bottom flask was equipped with an overhead stirrer,thermowell/thermometer, addition funnel (1 L), and cooling vessel(empty). The flask was flushed with nitrogen, charged with formic acid(877 g) and cooled with an ice bath. Triethylamine (755 g) was chargedto the addition funnel and added slowly over 50 min. to the stirredformic acid. After the addition was complete, the cooling bath wasremoved and the reaction solution was diluted with DMF (5.0 L). Theketoester (2648 g) was added in one portion, followed by an additional0.5 L of DMF. The flask was equipped with a heating mantle and thestirred mixture was heated gradually to 16° C. to dissolve all solids.The catalyst (S,S)-Ts-DPEN-Ru—Cl-(p-cymene) (18.8 g) was added in oneportion and the stirred mixture was heated to 55° C. over 1 h. Theresulting dark solution was stirred at 55° C. for 16 h. TLC indicatedthe reaction was complete. The solvent was evaporated under reducedpressure (Buchi R152 rotary evaporator under high vacuum, bath temp=60°C.) to give 3574 g of a brown oil. The oil was dissolved indichloromethane (10 L) and transferred to a 5 gal. stationary separatoryfunnel. The dark solution was washed with saturated sodium bicarbonatesolution (3.0 L) and the aqueous phase was back extracted withdichloromethane (3.0 L). The combined dichloromethane extracts weredried over MgSO₄ (300 g), filtered, and concentrated under reducedpressure to provide 3362 g of a brown oil.

Column: Chiralpak AD, 0.46×25 cm; mobile phase=10:90, ethanol:hexane,isocratic; flow rate=1.5 mL/min; injection volume=10 μL UV detection at254 nm.

Retention times: R-hydroxy ester=19.9 min.

-   -   S-hydroxy ester=21.7 min.

Retention times: R-diol=14.2 min.

-   -   S-diol=15.5 min

Hydroxy Ester:

¹H NMR (CDCl₃): δ 2.73 (d, 2H, J=1.5 Hz), 3.73 (s, 3H), 4.35 (s, 1H),5.11-5.19 (m, 1H), 7.31 (d, 2H, J=6.6 Hz), 8.53 (d, 2H, J=6.0 Hz).

Merck silica gel 60 plates, 2.5×7.5 cm, 250 micron; UV lamp: 5% MeOH inCH₂Cl₂; Rf of S.M.=0.44, Rf of product=0.15.

e.e.=87% S isomer of hydroxy ester.

Step C: Synthesis of S-(−)-1-(Pyrid-4-yl)-1,3-propanediol

A 22 L, 4-neck round bottom flask was equipped with an overhead stirrer,thermowell/thermometer, addition funnel (2 L), condenser and coolingvessel (empty). The flask was flushed with nitrogen and chargedsequentially with sodium borohydride (467 g, 12.3 mol), 1-butanol (9.0L), and water (148 mL, 8.23 mol) The crude hydroxyester was dissolved in1-butanol (1.0 L) and the solution was charged to the addition funnel.The solution was added over 3.25 h, using cooling as necessary to keepthe temperature below 62° C. After addition was complete, the mixturewas stirred for 0.5 h then the flask was equipped with a heating mantleand the stirred mixture was heated to 90° C. over 0.75 h. The mixturewas stirred at 90-93° C. for 2.25 h, then cooled over 1.5 h to 28° C.The reaction mixture was quenched with aqueous potassium carbonatesolution (10 wt/vol %, 6 L) and the mixture was stirred for 10 min. Thelayers were separated and the butanol phase was washed with aqueouspotassium carbonate solution (10 wt/vol %, 2 L) and sodium chloridesolution (15 wt/vol %, 2 L). The solvent was removed under reducedpressure (Buchi R152 rotary evaporator, high vacuum, bathtemperature=60° C.) until a concentrated solution resulted and 10.5 L ofdistillate had been collected. Acetonitrile (3 L) was fed into theevaporator flask and the solvent was evaporated under reduced pressure.Acetonitrile (9 L) was again fed into the evaporator flask and theslurry was stirred (rotation on the rotary evaporator) at ˜60° C. (bathtemperature=70° C., atmospheric pressure) for 15 min. The hot slurry wasfiltered through Celite 521 (250 g as a slurry in 1 L of acetonitrilewas prepacked on a 24 cm Buchner funnel). The filtrate was partiallyconcentrated under reduced pressure (5 L of distillate were collected)and the resulting slurry was heated at atmospheric pressure on therotary evaporator to dissolve all solids (bath temp=65° C.). The heatsource was turned off and the resulting solution was stirred on therotary evaporator for 10 h, with gradual cooling to ambient temperature.The resulting mixture was filtered and the collected solid was washedwith acetonitrile (2×200 mL) and dried to constant weight (−30 in. Hg,55° C., 4 h), giving S-(−)-1-(4-pyridyl)-1,3-propanediol as a yellowsolid weighing 496 g.

Melting point=98-100° C.

HPLC Conditions:

Column: Chiralpak AD, 0.46×25 cm; mobile phase=10:90, ethanol:hexane,isocratic; flow rate=1.5 mL/min; injection volume=10 μL UV detection at254 nm.

Retention times: R-diol=14.2 min.

-   -   S-diol=15.5 min.

Merck silica gel 60 plates, 2.5×7.5 cm, 250 micron; UV lamp; 15% MeOH inCH₂Cl₂; Rf of starting material=0.38, Rf of product=0.17, Rf of boroncomplex=0.26.

Example 9 Synthesis of (S)-3-(3′-chlorophenyl)-1,3-dihydroxypropane via(−)-β-chlorodiisopinocampheylborane (DIPCI) Reduction

Step A: Preparation of 3-(3-chlorophenyl)-3-oxo-propanoic acid:

A 12 L, 3-neck round bottom flask was equipped with a mechanical stirrerand addition funnel (2 L). The flask was flushed with nitrogen andcharged with diisopropylamine (636 mL) and THF (1.80 L). A thermocoupleprobe was immersed in the reaction solution and the stirred contentswere cooled to −20° C. n-Butyllithium (1.81 L of a 2.5 M solution inhexanes) was charged to the addition funnel and added slowly withstirring, maintaining the temperature between −20 and −28° C. After theaddition was complete (30 min), the addition funnel was rinsed withhexanes (30 mL) and the stirred solution was cooled to −62° C.Trimethylsilyl acetate (300 g) was added slowly with stirring,maintaining the temperature <−60° C. After the addition was complete (30min), the solution was stirred at −60° C. for 15 min. 3-Chlorobenzoylchloride (295 mL) was added slowly with stirring, maintaining thetemperature <−60° C. After the addition was complete (65 min), thecooling bath was removed and the reaction solution was stirred for 1.25h, with gradual warming to 0° C. The reaction flask was cooled with anice bath, then water (1.8 L) was added to the stirred solution. Thereaction mixture was stirred for 10 min., then diluted with t-butylmethyl ether (1.0 L). The lower aqueous phase was separated andtransferred to a 12 L, 3-neck round bottom flask equipped with amechanical stirrer. t-Butyl methyl ether was added (1.8 L) and thestirred mixture was cooled to <10° C. (ice bath). Concentrated HClsolution (300 mL of 12 M solution) was added and the mixture wasvigorously stirred. The layers were separated and aqueous phase wasfurther acidified with con. HCl (30 mL) and extracted again with t-butylmethyl ether (1.0 L). The combined MTBE extracts were washed with brine(1 L), dried (MgSO₄, 70 g), filtered and concentrated under reducedpressure to give 827 g of a yellow solid. The crude solid was slurriedin hexanes (2.2 L) and transferred to a 5 L, 3-neck round bottom flaskequipped with a mechanical stirrer. The mixture was stirred at <10° C.(ice bath) for 1 h, then filtered, washed with hexanes (4×100 mL) anddried to constant weight (−30 in. Hg, ambient temperature, 14 h).Recovery=309 g of a pale yellow powder.

Step B: Preparation of (S)-3-(3-chlorophenyl)-3-hydroxypropanoic Acid:

A 12 L, 3-neck round bottom flask was equipped with a mechanical stirrerand addition funnel (1 L). The flask was flushed with nitrogen andcharged with 3-(3-chlorophenyl)-3-oxo-propanoic acid (275.5 g) anddichloromethane (2.2 L). A thermocouple probe was immersed in thereaction slurry and the stirred contents were cooled to −20° C.Triethylamine (211 mL) was added over 5 min. to the stirred slurry andall solids dissolved. A dichloromethane solution of(−)-β-chlorodiisopinocampheylborate (1.60 M, 1.04 L) was charged to theaddition funnel, then added slowly with stirring, maintaining thetemperature between −20 and −25° C. After the addition was complete (35min), the solution was warmed to ice bath temperature (2-3° C.) andstirred for 4 h. Water (1.2 L) was added to the cloudy orange reactionmixture, followed by 3 M NaOH solution (1.44 L). The mixture wasvigorously stirred for 5 min, then transferred to a separatory funnel.The layers were separated and the basic aqueous phase was washed withethyl acetate (1.0 L). The aqueous phase was acidified with conc. HCl(300 mL) and extracted with ethyl acetate (2×1.3 L). The two acidicethyl acetate extracts were combined, washed with brine (600 mL), dried(MgSO₄, 130 g), filtered and concentrated under reduced pressure toprovide 328 g of a yellow oil (the oil crystallized on standing). Thesolid was slurried in ethyl acetate (180 mL) and transferred to a 2 L,3-neck round bottom flask, equipped with a mechanical stirrer. Thestirred mixture was cooled to <10° C. (ice bath), then diluted withhexanes (800 mL). The resulting mixture was stirred at ice bathtemperature for 4 h, then filtered. The collected solid was washed with4:1 hexanes:ethyl acetate (3×50 mL) and dried to constant weight (−30in. Hg, ambient temperature, 12 h). Recovery=207.5 g of a white powder.

Step C: Preparation of (S)-(−)-1-(3-chlorophenyl)-1,3-propanediol:

The compound was prepared as described in Example 7, Step D.

The residue was dissolved in methanol (1 mL) and analyzed by chiral HPLC(see, Example 7; Step B). ee>98%.

Example 10 The Preparation of 1,3-Diols via Catalytic AsymmetricHydrogenation Step A:

Beta-ketoester starting material was synthesized as described in Example7, step A.

Step B:

A solution containing beta-ketoester (1 mmol) in either methanol orethanol (5-10 mL/mmol ketoester) was degassed through several pump/vent(N₂) cycles at room temperature. The degassed solution was moved into aglove bag and under an atmosphere of N₂ was poured into a stainlesssteel bomb containing a stir bar and 1.0 mole % Ru-BINAP catalyst. Thebomb was sealed, removed from the glove bag and purged with H₂ prior tostirring 18-24 h at room temperature and 150 psi H₂. After venting thehydrogen pressure, the bomb was opened and the reaction mixture wasremoved and concentrated. The crude beta-hydroxyester was used forhydrolysis.

Step C:

Crude beta-hydroxy ester was hydrolyzed as described in Example 7, stepC.

Step D:

Optically active beta-hydroxy acid was reduced as described in Example7, step D.

Synthesis of Racemic Phosphorylating Agents: Example 11 GeneralProcedure for the Synthesis oftrans-4-(aryl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanes

Example 11.1 Synthesis oftrans-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

A solution of 1-(3-chlorophenyl)-1,3-propane diol (25 g, 134 mmol) andtriethylamine (62.5 mL, 4421 mmol) in THF was added to a solution of4-nitrophenyl-phosphorodichloridate (37.7 g, 147 mmol) in THE at roomtemperature and the resulting solution was heated at reflux. After 2 h,TLC indicated complete consumption of the starting diol and formation ofthe cis and trans isomers in a 60/40 ratio (HPLC). The clear yellowsolution was cooled to 30° C., sodium 4-nitrophenoxide (56 g, 402 mmol)was added and the reaction mixture was heated at reflux. After 90 min.the reddish reaction mixture was cooled to room temperature and stirredat room temperature for 2 h then placed in the refrigerator overnight.The final ratio was determined by HPLC to be 96/4 trans/cis. Thereaction mixture was quenched with a saturated solution of ammoniumchloride and diluted with ethyl acetate. The layers were separated andthe organics were washed 4 times with 0.3 N sodium hydroxide to removethe nitrophenol, then saturated sodium chloride and dried over sodiumsulfate. The filtered solution was concentrated under reduced pressureand the resulting solid was recrystallized from ethyl acetate to givelarge off white needles (45 g, mp=115-116° C., purity 98 A %).

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: cis-isomer 5.6-5.8 (m,1H), trans-isomer 5.5-5.6 9 (m, 1H).

TLC conditions: Merck silica gel 60 F254 plates, 250 μm thickness;mobile phase=60/40 hexanes/ethyl acetate; Rf: diol=0.1,cis-phosphate=0.2, trans-phosphate=0.35.

HPLC conditions: Column=Waters 1 Bondapack C18 3.9×300 mm; mobilephase=40/60 acetonitrile/phosphate buffer pH 6.2; flow rate=1.4 ml/min;detection=UV@270 nm; retention times in min: cis-isomer=14.46,trans-isomer=16.66, 4-nitrophenol=4.14.

Example 11.2 Synthesis oftrans-4-(3-pyrid-3-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(3-pyridyl)-1,3-propanediol.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.6-5.8(m, 1H)

Example 11.3 Synthesis oftrans-4-(3,-5-difluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3,-5-difluorophenyl)-1,3-propanediol. TLC conditions: Merck silicagel 60 F254 plates, 250 μm thickness; mobile phase=50/50 hexanes/ethylacetate; Rf: diol=0.1, cis-phosphate=0.25, trans-phosphate=0.4.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.7-5.5(m, 1H)

Example 11.4 Synthesis oftrans-4-(4-methylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(4-methylphenyl)-1,3-propanediolTLC: 50/50 hexanes/ethyl acetate; Rf: cis-phosphate=0.25;trans-phosphate=0.35.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.65-5.5(m, 1H)

Example 11.5 Synthesis oftrans-4-(3,5-dimethylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3,5-dimethylphenyl)-1,3-propanediol TLC: 50/50 hexanes/ethyl acetate;Rf: cis-phosphate=0.2; trans-phosphate=0.3.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.6-5.45(m, 1H)

Example 11.6 Synthesis oftrans-4-(3,5-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3,5-dichlorophenyl)-1,3-propanediol TLC: 70/30 hexanes/ethyl acetate;Rf: cis-phosphate=0.3; trans-phosphate=0.5.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.85-5.7(m, 1H)

Example 11.7 Synthesis oftrans-4-(pyrid-4-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(pyrid-4-yl)-1,3-propanediol TLC:95/5 dichloromethane/ethanol; Rf: trans-phosphate=0.35.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.7-5.55(m, 1H)

Example 11.8 Synthesis oftrans-4-(3-methoxycarbonylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3-methoxycarbonylphenyl)-1,3-propanediol TLC: 30/70 hexanes/ethylacetate; Rf: cis-phosphate=0.5; trans-phosphate=0.6.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.7-5.6(m, 1H)

Example 11.9 Synthesis oftrans-4-(4-methoxycarbonylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(4-methoxycarbonylphenyl)-1,3-propanediol TLC: 30/70 hexanes/ethylacetate; Rf: cis-phosphate=0.35; trans-phosphate=0.5.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.7-5.6(m, 1H)

Example 11.10 Synthesis oftrans-4-(5-bromopyrid-3-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(5-bromopyrid-3-yl)-1,3-propanediol

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.8-5.65(m, 1H)

Example 11.11 Synthesis oftrans-4-(2,3-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2,3-dichlorophenyl)-1,3-propanediol except that the isomerization wasconducted with 4-nitrophenol and lithium hydride as in Example 13a.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 6-5.9 (m,1H)

Example 11.12 Synthesis oftrans-4(2,3,5-trichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2,3,5-trichlorophenyl)-1,3-propanediol except that the isomerizationwas conducted with 4-nitrophenol and triethyl amine as in Example 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.9-5.7(m, 1H)

Example 11.13 Synthesis oftrans-4-(2-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(2-chlorophenyl)-1,3-propanediolexcept that the isomerization was conducted with 4-nitrophenol andlithium hydride as in Example 13a.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 6-5.9 (m,1H)

Example 11.14 Synthesis oftrans-4-(3,5-dimethoxyphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3,5-dimethoxyphenyl)-1,3-propanediol except that the isomerizationwas conducted with 4-nitrophenol and triethylamine as in Example 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.55-5.45(m, 1H), 3.3 (s, 6H)

Example 11.15 Synthesis oftrans-4-(2-bromophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(2-bromophenyl)-1,3-propanediolexcept that the isomerization was conducted with 4-nitrophenol andtriethylamine as in Example 13a.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.95-5.85(m, 1H)

Example 11.16 Synthesis oftrans-4-(3-bromo-5-ethoxyphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3-bromo-5-ethoxyphenyl)-1,3-propanediol except that the isomerizationwas conducted with 4-nitrophenol and triethylamine as in Example 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.9-5.75(m, 1H), 4.04 (q, 2H), 1.39 (t, 3H).

Example 11.17 Synthesis oftrans-4-(2-trifluoromethylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2-trifluoromethylphenyl)-1,3-propanediol except that theisomerization was conducted with 4-nitrophenol and triethylamine as inExample 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 6-5.75(m, 1H).

Example 11.18 Synthesis oftrans-4-(4-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(4-chlorophenyl)-1,3-propanediolexcept that the trans-isomer was isolated from the cis/trans mixturewithout isomerization. TLC: hexanes/ethyl acetate 1/1; Rf:cis-phosphate=0.2; trans-phosphate=0.6.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.6-5.5(m, 1H).

Example 11.19 Synthesis oftrans-4-(3-methylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(3-methylphenyl)-1,3-propanediolexcept that the trans-isomer was isolated from the cis/trans mixturewithout isomerization. TLC: hexanes/ethyl acetate 6/4; Rf:cis-phosphate=0.2; trans-phosphate=0.5.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.65-5.5(m, 1H).

Example 11.20 Synthesis oftrans-4-(4-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanes

Same as Example 11.1 starting with 1-(4-fluorophenyl)-1,3-propanediolexcept that the trans-isomer was isolated from the cis/trans mixturewithout isomerization.

¹H NMR (DMSO-d₆, Varian Gemini 200 MHz): C′-proton: trans-isomer5.78-5.85 (m, 1H).

Example 11.21 Synthesis oftrans-4-(2-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(2-fluorophenyl)-1,3-propanediolexcept that the trans-isomer was isolated from the cis/trans mixturewithout isomerization.

¹H NMR (DMSO-d₆, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.9-6.1(m, 1H).

Example 11.22 Synthesis oftrans-4-(3-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(3-fluorophenyl)-1,3-propanediolexcept that the trans-isomer was isolated from the cis/trans mixturewithout isomerization.

¹H NMR (DMSO-d₆, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.8-5.9(m, 1H).

Example 11.23 Synthesis oftrans-4-[4-(4-chlorophenoxy)phenyl]-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-[4-(4-chlorophenoxy)phenyl]-1,3-propanediol except that thetrans-isomer was isolated from the cis/trans mixture withoutisomerization.

¹H NMR (DMSO-d₆, Varian Gemini 200 MHz): C′-proton: trans-isomer5.75-5.9 (m, 1H).

Example 11.24 Synthesis oftrans-4-(3-bromophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(3-bromophenyl)-1,3-propanediolexcept that the trans-isomer was isolated from the cis/trans mixturewithout isomerization. TLC: hexanes/ethyl acetate 1/1; Rf:cis-phosphate=0.25; trans-phosphate=0.5.

¹H NMR (DMSO-d₆, Varian Gemini 200 MHz): C′-proton: trans-isomer5.8-5.95 (m, 1H).

Example 11.25 Synthesis oftrans-4-(3,4-ethylenedioxyphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3,4-ethylenedioxyphenyl)-1,3-propanediol except that the trans-isomerwas isolated from the cis/trans mixture without isomerization. TLC:hexanes/ethyl acetate 1/1; Rf: trans-phosphate=0.6.

¹H NMR (DMSO-d₆, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.8-5.9(m, 1H).

Example 11.26 Synthesis oftrans-4-(2-fluoro-4-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2-fluoro-4-chlorophenyl)-1,3-propanediol except that the trans-isomerwas isolated from the cis/trans mixture without isomerization. TLC:hexanes/ethyl acetate 1/1; Rf: trans-phosphate=0.7.

¹H NMR (DMSO-d₆, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.9-6(m, 1H).

Example 11.27 Synthesis oftrans-4-(2,6-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2,6-dichlorophenyl)-1,3-propanediol except that the trans-isomer wasisolated from the cis/trans mixture without isomerization. TLC:hexanes/ethyl acetate 1/1; Rf: trans-phosphate=0.65.

¹H NMR (DMSO-d₆, Varian Gemini 200 MHz): C′-proton: trans-isomer 6.2-6.4(m, 1H).

Example 11.28 Synthesis oftrans-4-(2-fluoro-5-methoxyphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2-fluoro-5-methoxyphenyl)-1,3-propanediol except that thetrans-isomer was isolated from the cis/trans mixture withoutisomerization.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.8-5.95(m, 1H), 3.8 (s, 3H).

Example 11.29 Synthesis oftrans-4-(3-fluoro-4-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3-fluoro-4-chlorophenyl)-1,3-propanediol except that theisomerization was conducted with 4-nitrophenol and triethylamine as inExample 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.4-5.6(m, 1H).

Example 11.30 Synthesis oftrans-4-(3-chloro-4-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3-chloro-4-fluorophenyl)-1,3-propanediol except that theisomerization was conducted with 4-nitrophenol and triethylamine as inExample 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.5-5.6(m, 1H).

Example 11.31 Synthesis oftrans-4-(2-fluoro-5-bromophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2-fluoro-5-bromophenyl)-1,3-propanediol except that the isomerizationwas conducted with 4-nitrophenol and triethylamine as in Example 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.8-5.9(m, 1H).

Example 11.32 Synthesis oftrans-4-(2,3,5,6-tetrafluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2,3,5,6-tetrafluorophenyl)-1,3-propanediol except that theisomerization was conducted with 4-nitrophenol and triethylamine as inExample 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.9-6 (m,1H).

Example 11.33 Synthesis oftrans-4-(2,3,6-trifluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2,3,6-trifluorophenyl)-1,3-propanediol except that the isomerizationwas conducted with 4-nitrophenol and triethylamine as in Example 13b.

¹H NMR (CDCl₃, Varian Gemini 200 MHz): C′-proton: trans-isomer 5.9-6 (m,1H).

Example 11.34 Synthesis oftrans-4(R)-(phenyl)-2-(4-chlorophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with l(R)-(phenyl)-1,3-propanediolisolated by column without the isomerization.

Rf=0.5 (50% EtOAc in Hexanes). mp 90-92° C. Anal calcd for C₁₅H₁₄ClO₄P:C, 55.49; H, 4.35. Found: C, 55.64; H, 3.94.

Example 11.35 Synthesis oftrans-4(R)-(phenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with l(R)-(phenyl)-1,3-propanediolisolated by column without the isomerization.

Rf=0.4 (50% EtOAc in Hexanes). mp 130-131° C. Anal calcd for C₁₅H₁₄NO₆P:C, 53.74; H, 4.21; N, 4.18. Found: C, 53.86; H, 4.07; N, 4.00.

Example 11.36 Synthesis oftrans-4(S)-(phenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with l(S)-(phenyl)-1,3-propanediol.

Rf=0.2 (5% EtOAc in CH₂Cl₂). mp 128-129° C. Anal calcd for C₁₅H₁₄NO₆P:C, 53.74; H, 4.21; N, 4.18. Found: C, 53.69; H, 4.53; N, 4.23.

Example 11.37 Synthesis oftrans-4-(3-trifluoromethylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3-trifluoromethylphenyl)-1,3-propanediol.

Rf=0.32(35% EtOAc in hexanes). mp 78-81° C. Anal calcd for C₁₆H₁₃F₃NO₆P:C, 47.66; H, 3.25; N, 3.47. Found: C, 47.69; H, 3.77; N, 3.52.

Example 11.38 Synthesis oftrans-4-(2,4-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2,4-dichlorophenyl)-1,3-propanediol.

Rf=0.32(35% EtOAc in hexanes). mp 154-157° C. Anal calcd forC₁₅H₁₂C₁₂NO₆P: C, 44.58; H, 2.99; N, 3.47. Found: C, 44.63; H, 3.07; N,3.47.

Example 11.39 Synthesis oftrans-4-(3-bromo-4-fluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3-bromo-4-fluorophenyl)-1,3-propanediol. Rf=0.2 (5% EtOAc in CH₂Cl₂).mp 108° C. Anal calcd for C₁₅H₁₂NO₆BrFP: C, 41.69; H, 2.80; N, 3.24.Found: C, 41.90; H, 2.76; N, 3.05.

Example 11.40 Synthesis oftrans-4-(2-pyridyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(2-pyridyl)-1,3-propanediol. mp99-102° C. Anal calcd for C₁₄H₁₃N₂O₆P: C, 50.01; H, 3.90; N, 8.33.Found: C, 49.84; H, 3.41; N, 8.14.

Example 11.41 Synthesis oftrans-4-(3,4-dichlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(3,4-dichlorophenyl)-1,3-propanediol. Rf=0.15 (35% EtOAc in Hexanes).mp 126-129° C. Anal calcd for C₁₅H₁₂C₁₂NO₆P: C, 44.58; H, 2.99; N, 3.47.Found: C, 44.71; H, 3.49; N, 3.41.

Example 11.42 Synthesis oftrans-4-(4-tert-butylphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(4-tert-butylphenyl)-1,3-propanediol. Rf=0.20 (35% EtOAc in Hexanes).mp 108-111° C. Anal calcd for Cl₉H₂₂NO₆P: C, 58.31; H, 5.67; N, 3.58.Found: C, 58.04; H, 5.67; N, 3.55.

Example 11.43 Synthesis oftrans-4-(3-thiophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(3-thiophenyl)-1,3-propanediol. mp94-96° C. Anal calcd for C₁₃H₁₂NO₆PS: C, 45.75; H, 3.54; N, 4.10. Found:C, 45.65; H, 3.21; N, 4.24.

Example 11.44 Synthesis oftrans-4-(3-furanyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with 1-(3-furanyl)-1,3-propanediol. mp108-111° C. Anal calcd for C₁₃H₁₂NO₇P: C, 48.01; H, 3.72; N, 4.31.Found: C, 48.06; H, 3.61; N, 4.26.

Example 11.45 Synthesis oftrans-4-(2-bromo-5-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2-bromo-5-chlorophenyl)-1,3-propanediol. Rf=0.20 (5% MeOH in CH₂Cl₂).mp 105-106° C. Anal calcd for C₁₅H₁₂NO₆BrClP: C, 40.16; H, 2.70; N,3.12. Found: C, 39.97; H, 2.86; N, 3.06.

Example 11.46 Synthesis oftrans-4-(2,5-difluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2,5-difluorophenyl)-1,3-propanediol. Rf=0.50 (50% EtOAc in Hexanes).mp 120-122° C. Anal calcd for C15H₁₂F₂NO₆P: C, 48.53; H, 3.26; N, 3.77.Found: C, 48.46; H, 3.52; N, 3.87.

Example 11.47 Synthesis oftrans-4-(2,4-difluorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with1-(2,4-difluorophenyl)-1,3-propanediol. Rf 0.50 (50% EtOAc in Hexanes).mp 85-87° C. Anal calcd for C₁₅H₁₂F₂NO₆P: C, 48.53; H, 3.26; N, 3.77.Found: C, 48.82; H, 3.55; N, 3.84.

Example 11.48 Synthesis oftrans-4-cis-6-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with trans-1,3-diphenyl-1,3-propanediol(Yamamura, H., Araki, S., Tetrahedron 53: 46 15685-15690 (1997)) withoutequilibration. Rf=0.29 (35% EtOAc in Hexanes). mp 118-121° C. Anal calcdfor C₂₁H₁₈NO₆P: C, 61.32; H, 4.41; N, 3.41. Found: C, 60.94; H, 4.44; N,3.53.

Example 11.49 Synthesis oftrans-4-tranis-6-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with cis-1,3-diphenyl-1,3-propanediol(Yamamura, H., Araki, S., Tetrahedron 53: 46 15685-15690 (1997)) withoutequilibration. Rf=0.65 (5% EtOAc in CH₂Cl₂). mp 144-147° C. Anal calcdfor C₂₁H₁₈NO₆P: C, 61.32; H, 4.41; N, 3.41. Found: C, 61.21; H, 4.58; N,3.36.

Example 11.50 Synthesis ofcis-4-cis-6-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with cis-1,3-diphenyl-1,3-propanediol(Yamamura, H., Araki, S., Tetrahedron 53: 46, 15685-15690 (1997))without equilibration. Rf=0.3 (5% EtOAc in CH₂Cl₂). mp 135-138° C. Analcalcd for C₂₁H₁₈NO₆P: C, 61.32; H, 4.41; N, 3.41. Found: C, 61.29; H,4.77; N, 3.46.

Example 11.51 Synthesis ofcis-4-cis-5-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with cis-1,2-diphenyl-1,3-propanediol(Kristersson, P, Lindquist, K., Acta Chem. Scand. B 34:213-234 (1980))without equilibration. Rf=0.35 (5% EtOAc in CH₂Cl₂). mp 136-139° C. Analcalcd for C₂₁H₁₈NO₆P: C, 61.32; H, 4.41; N, 3.41. Found: C, 60.95; H,4.41; N, 3.82.

Example 11.52 Synthesis oftrans-4-trans-5-(diphenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with cis-1,2-diphenyl-1,3-propanediol(Kristersson, P, Lindquist, K., Acta Chem. Scand. B 34:213-234 (1980))without equilibration. Rf=0.65 (5% EtOAc in CH₂Cl₂). mp 176-178° C. Analcalcd for C₂₁H₁₈NO₆P: C, 61.32; H, 4.41; N, 3.41. Found: C, 61.09; H,4.46; N, 3.80.

Example 11.53 Synthesis oftrans-4,4-dimethyl-6-(phenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinaneStep A:

To a solution of diisopropylamine (58.4 g, 577 mmol) in dry ether (500mL) at −78° C. under nitrogen was added n-BuLi (215 mL, 2.5 M in hexane,538 mmol) over 30 min. The reaction was stirred for 10 min beforeaddition of ethyl acetate (55 mL, 558 mmol) over a period 30 min.Freshly distilled benzaldehyde (47 mL, 443 mmol) in ether (50 mL) wasslowly added over 30 min and the mixture was allowed to warm to roomtemperature. The reaction was quenched with saturated ammonium chloride(150 mL) at 0° C. The organic layer was washed, dried (anhydrous Na₂SO₄)and concentrated to give the crude addition product.

Step B:

To a solution of crude condensation product (10.6 g, 54.6 mmol) in dryether at −78° C. was added MeMgBr (60 mL, 3.0 M in THF, 180 mmol). Themixture was allowed to warm to room temperature and stirred overnight.The reaction was quenched with ammonium chloride (50 mL) at 0° C. anddiluted with EtOAc (350 mL). The organic layer was washed, dried(anhydrous Na₂SO₄) and concentrated. The crude product was purified bycolumn chromatography (0-10% EtOAc in CH₂Cl₂) to give3,3-dimethyl-1-phenyl-1,3-propanediol (7 g) as a pale yellow oil.

Step C:

Same as Example 11.1 starting with 3,3-dimethyl-1-phenyl-1,3-propanediolwithout equilibration. Rf=0.18 (35% EtOAc in hexanes). mp 131-133° C.Anal calcd for C₁₇H₁₈NO₆P: C, 56.20; H, 4.99; N, 3.86. Found: C, 56.00;H, 5.03; N, 3.86.

Example 11.54 Synthesis ofcis-4-(3-chlorophenyl)-cis-5-methoxy-(2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinaneandtrans-4-(3-chlorophenyl)-cis-5-methoxy-(2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane(Example 11.55) Step A:

To a solution of lithium diisopropylamide (356 mmol) in THF (500 mL) at−78° C. was slowly added 2-methoxy-methyl acetate (38.8 mL, 392 mmol)via an addition funnel. The reaction was stirred at −78° C. for 30 minbefore 3-chlorobenzaldehyde (20.1 mL, 178 mmol) was added. The reactionwas allowed to warm to room temperature and quenched with saturated aqNH₄Cl (500 mL). The mixture was extracted with EtOAc (3×200 mL) and thecombined organic extracts were washed with water and dried (anhydrousNa₂SO₄). The crude product was purified by column chromatography (5-50%EtOAc in hexanes) to yield 3-(3-chlorophenyl)-3-hydroxy-2-methoxy-methylproprionate (39 g) as pale yellow oil.

Step B:

To a solution of the ester (39 g, 159 mmol) obtained from step A inethanol (500 mL) was added sodium borohydride (6.2 g, 159 mmol) in threeportions, over 10 min. The reaction was refluxed for 3 h and the ethanolwas evaporated under reduced pressure. The residue was dissolved inEtOAc (500 mL), washed with water and dried (anhydrous Na₂SO₄). Thecrude product was purified by column chromatography (1-5% MeOH—CH₂Cl₂)to give the diol (28 g) as colorless oil.

Step C:

To a solution of diol (28 g, 129 mmol) in acetone (250 mL) was addedtrimethyl orthoformate (10 mL) followed by p-toluenesulfonic acid (500mg, 2.64 mmol) and the reaction was heated to reflux overnight. Thereaction was cooled to room temperature and the acetone was removedunder vacuum. The residue was dissolved in ethyl acetate and washed withNaHCO₃, water and dried (anhydrous Na₂SO₄). The ketals were separated bycolumn chromatography (5-10% EtOAc in hexanes) to give 1,2-cis (7.26 g)and 1,2-trans ketal (0.9 g) diastereomers.

Step D:

The 1,2-cis ketal (4.5 g, 17.5 mmol) was dissolved in 70% aq TFA (10 mL)and allowed to react overnight at room temperature. The reaction wasdiluted with acetonitrile (30 mL) and volatiles were removed underreduced pressure. The residue was dissolved in EtOAc (300 mL) and theorganic layer was washed with saturated aq NaHCO₃, water and dried(anhydrous Na₂SO₄). The crude product was purified by columnchromatography (1-5% MeOH—CH₂Cl₂) to give 1,2-cis diol diastereomer (3.5g). The 1,2-trans ketal diastereomer was also hydrolyzed following theabove procedure to give 1,2-trans-diol diastereomer.

Step E:

1,2-cis-diol diastereomer was subjected to phosphorylation using theprocedure described in Example 11.1 without equilibration to give thefollowing two isomers.

Example 11.54 Rf=0.57 (5% EtOAc in CH₂Cl₂). mp 110-112° C.

Anal calcd for C₁₆H₁₅NO₇PCl: C, 48.08; H, 3.78; N, 3.50. Found: C,48.35; H, 3.56; N, 3.69.

Example 11.55 Rf=0.34 (5% EtOAc in CH₂Cl₂). mp 131-134° C.

Anal calcd for C₁₆H₁₅NO₇PCl.0.3H₂O: C, 47.44; H, 3.88; N, 3.46. Found:C, 47.23; H, 4.01; N, 3.46.

Example 12 General Procedure for the Synthesis oftrans-4-(aryl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanes UsingPhosphorus Oxychloride

Phosphorus oxychloride (3.4 mL, 36.3 mmol) was added to a solution of1-(3-chlorophenyl)-1,3-propanediol in dichloromethane at 0° C. followedby triethylamine (10.2 mL, 73 mmol). After 2 h, sodium 4-nitrophenoxide(10.63 g, 66 mmol) was added to the solution of cis/transphosphorochloridate reagent and the orange reaction mixture was heatedat reflux for 1 h. The cooled solution was partitioned with ethylacetate and a saturated solution of ammonium chloride. The organics wereseparated and dried over sodium sulfate, filtered and concentrated underreduced pressure. The residue was taken up in THF, sodium4-nitrophenoxide (10.63 g, 66 mmol) was added and the orange reactionmixture was heated to reflux for 3 h (IPLC, 95/5 trans/cis). The cooledsolution was partitioned with ethyl acetate and a saturated solution ofammonium chloride. The organics were separated and washed with 0.3 Nsolution of sodium hydroxide and brine, dried over sodium sulfate andconcentrated under reduced pressure. Recrystallization from ethylacetate as in Example 10 gave the phosphate reagent.

Example 13 Procedures for the Enrichment in trans-isomer of a cis/transMixture of 4-(aryl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

A cis/trans mixture of4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinaneswas prepared as in Example 11, except that the cis and trans isomerswere separated by column chromatography prior to the addition of4-nitrophenol.

cis-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanewas isomerized to the trans isomer by adding a solution of thecis-isomer to a solution of 4-nitrophenoxide prepared with the followingbases.

Example 13a

Lithium hydride (19.4 mg, 2.44 mmol) was added to a solution of4-nitrophenol in THF at room temperature. The yellow solution wasstirred at room temperature for 30 min. A solution ofcis-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane(300 mg, 0.813 mmol) in THF was added to the solution of lithium4-nitrophenoxide. The orange reaction mixture was stirred a roomtemperature. After 5 h the ratio was 92.9/5.4 trans/cis (HPLCdetermination).

Example 13b

Same as above using triethylamine instead of lithium hydride. After 20 hthe trans/cis ratio was 90.8/5.3.

Example 13c

Same as above using DBU instead of lithium hydride. After 3 h thetrans/cis ratio was 90.8/5.3.

Synthesis of Enantioenriched Phosphorylating Agents

Example 14 General Procedure for the Synthesis of Enantioenrichedtrans-4-(aryl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinanes

Example 14a Synthesis of(+)-(4R)-trans-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

A solution of (+)-(R)-1-(3-chlorophenyl)-1,3-propanediol (3 g, 16.1mmol) and triethylamine (6.03 ml, 59.6 mmol) in THF (80 mL) was addeddropwise to a solution of 4-nitrophenoxyphosphorodichloridate (7.63 g,29.8 mmol) in 150 mL of THF at 0° C. After about 2 h, the starting diolwas consumed, with the formation of two isomeric4-nitrophenylphosphates, and additional triethylamine (8.31 mL) followedby of 4-nitrophenol (8.29 g, 59.6 mmol) were added. The reaction mixturewas stirred overnight. The solvent was evaporated under reduced pressureand the residue was partitioned between ethyl acetate and water. Theorganic phase was washed (0.4 M NaOH, water and sat'd NaCl solution) anddried over MgSO₄. Concentration and chromatography of the residue using30% ethyl acetate in hexanes yielded 4.213 g of the desired product.

¹H NMR (200 MHz, CDCl₃): 8.26 (2H, d, J=9.7 Hz), 7.2-7.5 (6H, m), 5.56(1H, apparent d, J=11.7 Hz), 4.4-4.7 (2H, m), 2.2-2.6 (1H, m), 2.0-2.2(1H, m).

mp: 114-115° C. [α]_(D)=+91.71. Elemental Analysis: Calculated forC₁₅H₁₃NO₆ClP: C, 48.73; H, 3.54; N, 3.79. Found: C, 48.44; H, 3.20; N,3.65

Example 14b Synthesis of(−)-(4S)-trans-4-(3-chlorophenyl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

In a similar manner, from 3.116 g of(−)-(S)-1-(3-chlorophenyl)-1,3-propane diol was obtained 4.492 g of thedesired phosphate. ¹H NMR (200 MHz, CDCl₃): 8.26 (2H, d, J=9.7 Hz),7.2-7.5 (6H, m), 5.56 (1H, apparent d, J=11.7 Hz), 4.414.7 (2H, m),2.2-2.6 (1H, m), 2.0-2.2 (1H, m). mp: 114-115° C. [α]_(D)=−91.71.Elemental Analysis: Calculated for C₁₅H₁₃NO₆ClP: C, 48.73; H, 3.54, N,3.79. Found: C, 48.61; H, 3.36; N, 3.66.

Example 14c Synthesis of(−)-(4S)-trans-phenyl-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

Same as Example 11.1 starting with S-(−)-1-phenyl-1,3-propanediol exceptthat the isomerization was conducted with 4-nitrophenol andtriethylamine as in Example 13b.

TLC: hexanes/ethyl acetate 4/1); Rf=0.4

¹H NMR (DMSO-d₆, Varian Gemini 300 MHz): C′-proton: trans-isomer5.85-5.75 (m, 1H).

Example 15 General Procedures for Maintaining Enantiomeric Excess DuringSynthesis of Enantioenriched Phosphorylating Reagent Example 15aSynthesis of(−)-(4S)-trans-(pyrid-4-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

A 12 L round bottom flask equipped with an overhead stirrer and nitrogeninlet was charged with (S)-(−)-1-(pyrid-4-yl)-1,3-propanediol (1.2 kg,7.83 mol) and pyridine (6 L) The mixture was vigorously stirred at roomtemperature for 0.5 h until all the solids had dissolved. Meanwhile, a22 L, 3-neck flask was equipped with an overhead stirrer, thermocouple,cooling bath, and nitrogen inlet. This vessel was charged with4-nitrophenyl phosphorodichloridate (2.01 kg, 7.83 mol) and pyridine (6L). The resulting mixture was cooled to 3.3° C. After the diol wascompletely dissolved (0.5 h), triethylamine (190 mL, 1.36 mol) was addedand the slightly cloudy, yellow-brown solution was transferred inportions to a 2 L addition funnel on the 22 L flask. The solution wasadded to the cold phosphorodichloridate solution over 3.25 h. After theaddition was complete, the cooling bath was drainedand stirring wascontinued for 3 h. During this time, a 50 L, 3-neck flask was equippedwith an overhead stirrer, thermocouple, addition funnel, cooling bath(ice water) and nitrogen inlet. This flask was then charged with sodiumhydride (180 g, 4.5 mol) and THF (1 L) and the addition funnel wascharged with a solution of 4-nitrophenol (817 g, 5.87 mol) in THF (1 L).The nitrophenol solution was slowly added to the cold suspension ofsodium hydride. After the addition was complete, the resulting brightorange suspension was stirred at room temperature for 1 h. After thediol-dichloridate reaction was judged complete the dark suspension wassubjected to vacuum filtration. The glassware and filter cake(triethylamine-HCl) were rinsed with THF (1 L) and the combined filtrateand rinse were poured into the orange, sodium 4-nitrophenoxidesuspension. The resulting mixture was then heated at 40° C. for 3.5 h atwhich time the heating mantle was turned off and the reaction wasstirred an additional 11 h at room temperature. The crude reactionmixture was concentrated on a rotary evaporator at 45-50° C. at reducedpressure (oil pump). The resulting thick, black, foamy tar was dissolvedin 1.5 M aq HCl (12 L) and ethyl acetate (8 L). The mixture wastransferred to a 12.5-gallon separatory funnel, stirred 10 min, and thephases separated. The ethyl acetate layer was washed with an additional1.3 L of 1.5 M aq HCl. To the combined aqueous layers was addeddichloromethane (8 L) and the vigorously stirred mixture was carefullyneutralized with solid sodium bicarbonate. The layers were separated andthe aqueous layer was extracted with dichloromethane (8 L). The combinedorganic layers were dried over magnesium sulfate (600 g) and filtered.The solution was concentrated on a rotary evaporator until most of thesolvent was removed and a thick suspension resulted. 2-Propanol (5 L)was added and evaporation continued until 4 L of distillate werecollected. 2-Propanol (3 L) was added and evaporation continued until 3L of distillate were collected. The thick slurry was diluted with2-propanol (2 L) and the mixture stirred with cooling (ice bath) for 1h. The solid was collected by filtration, washed with 2-propanol (2 L),and dried in a vacuum oven (−30 in. Hg, 55° C., 18 h) to a constantweight of 1.86 kg. mp 140-142° C.

Specific Rotation=−80.35° (c=1.0, MeOH); ee=99+% trans.

HPLC Conditions:

Column: Chiralpak AD, 0.46×25 cm; mobile phase=50:50, 2-propanol:hexane,isocratic; flow rate=1.0 mL/min; injection volume=10 μL UV detection at254 nm.

The cis/trans equilibration was monitored by HPLC. Stopped at 92% trans,6.6% cis, r.t.=trans isomer 6.9 min. and cis isomer 10.9 min.

¹H NMR (DMSO-d6): δ=2.23-2.29 (m, 2H), 4.56-4.71 (m, 2H), 5.88-5.95 (m,1H), 7.44 (d, 2H, J=5.8 Hz), 7.59 (d, 2H, J=9.2 Hz), 8.34 (d, 2H, J=9.4Hz), 8.63 (d, 2H, J=5.8 Hz)

Example 15b Synthesis of(−)-(4S)-(−)-(pyrid-4-yl)-2-(4-nitrophenoxy)-2-oxo-1,3,2-dioxaphosphorinane

A 1 liter 3-neck round bottom flask was equipped with a mechanicalstirrer, addition funnel, a thermometer and a N₂ inlet. The flask ischarged with S-(−)-1-(pyrid-4-yl)-propane-1,3-diol (25 g, 163.4 mmol)and ethyl acetate (250 mL) and the resulting suspension was treatedslowly with a 4N HCl solution in dioxane (43 mL, 176 mmol) over a periodof 15 min. After stirring for 30 min at room temperature,4-nitrophenylphosphorodichloridate (41.81 g, 163.4 mmol) was added as asolid as quickly as possible under a positive flow of N₂. The internaltemperature of the reaction mixture was adjusted to −110° C. with thehelp of a dry ice-acetone cooling bath. A solution of triethylamine (79mL, 572 mmol) in ethyl acetate (100 mL) was added maintaining thereaction temperature at <−5° C. Thirty minutes after the completeaddition of the triethylamine solution, the cooling bath was removed andthe reaction mixture was stirred at room temperature for 1 h. Thereaction mixture was filtered to remove triethylamine-hydrochloridesalt, which is washed with ethyl acetate (3×30 mL) until the filtrateshows only faint absorption. The filtrate was evaporated down to avolume of 150-175 mL under reduced pressure. 4-Nitrophenol (7.5 g, 54.3mmol) and triethylamine (9 mL) were added to the concentrated solutionand the resulting orange reaction mixture was stirred at roomtemperature for 24 h. The solid formed in the reaction mixture wascollected by filtration, washed with ethyl acetate (2×25 mL) and methylt-butyl ether (25 mL) and dried under vacuum at 55° C. to give 31.96 gof the desired product. Same analytical data as Example 14a.

Example 16 Preparation of prodrugs of 2′-C-beta-methyl-7-deazaadenosinevia trans-phosphate Addition Example 16.14-Amino-7-(cis-5′-O-[4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

Step A:

To a solution of4-amino-7-(2-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(US2002-0147160A1, WO 02/057827) (10 g, 0.356 mol) in anhydrous acetone(145 mL) and anhydrous DMF (35 mL) were added p-toluene sulfonic acidmonohydrate (33.8 g, 0.18 moles) and triethyl orthoformate (31.2 mL,28.5 moles) at room temperature. The reaction was warmed to 80° C. andallowed to stir for 3 h under nitrogen. The mixture was evaporated underreduced pressure. The oily residue was purified by column chromatography(5% MeOH in CH₂Cl₂) to give the isopropylidene derivative (8.6 g) as awhite solid.

Step B:

To a solution of2′,3′-O-isopropylidene-4-amino-7-(2-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine(0.094 g, 0.29 mmol) in DMF (1.5 mL) was added t-butyl magnesiumchloride and stirred under nitrogen for 30 min. The reaction mixture wasthen cooled to −55° C. and the phosphorylating agent (whose preparationis described in example 11.1) (0.13 g, 0.35 mmol) in DMP (1.5 mL) wasadded dropwise. The reaction was allowed to warm to room temperature andstirred under nitrogen for 2 h. The mixture was evaporated under reducedpressure and purified by chromatography (5% MeOH in CH₂Cl₂) to yield0.070 g of the 2′,3′-O-isopropylidene protected prodrug as a yellowsolid.

Step C:

The prodrug (0.15 g, 0.27 mmol) obtained from the above step wasdissolved in pre-cooled 75% TFA/H₂O (20 mL) and allowed to stir at 0° C.for 2 h. The reaction mixture was evaporated under reduced pressure. Thecrude product was purified by flash chromatography (1% aq.NH₄OH in 10%MeOH in CH₂Cl₂) to give 0.142 g of the title compound as an off-whitesolid.

R_(f)=0.40 (10% MeOH in CH₂Cl₂). mp 138-141° C. Anal calcd forC₂₁H₂₄ClN₄O₇P.0.4 CH₂Cl₂: C, 47.18; H, 4.59; N, 10.28. Found: C, 46.97;H, 4.59; N, 10.11.

The following examples were synthesized as described in steps A-C ofexample 16.1, utilizing the phosphorylating agents of examples 1-15.

Example 16.24-Amino-7-(cis-5′-O-[4-(2,5-difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.35 (10% MeOH in CH₂Cl₂). mp 145-148° C. Anal Calcd forC₂₁H₂₃N₄O₇F₂P.1.35H₂O.1.0 CF₃CO₂H: C, 42.45; H, 4.14; N, 8.62. Found: C,42.18; H, 3.77; N, 8.42.

Example 16.34-Amino-7-(cis-5′-O-[4-(3-chloro-4-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.30 (10% MeOH in CH₂Cl₂). mp 128-130° C. Anal Calcd forC₂₁H₂₃N₄O₇FClP.2H₂O.1.9CF₃CO₂H: C, 38.11; H, 3.73; N, 7.17. Found: C,38.04; H, 3.28; N, 7.02.

Example 16.44-Amino-7-(cis-5′-O-[6,6-dimethyl-4-phenyl-2-oxo-1,3,2-dioxaphos-phorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.40 (10% MeOH in CH₂Cl₂). mp 140-142° C. Anal Calcd forC₂₃H₂₉N₄O₇P.1H₂O.0.4 CF₃CO₂H: C, 50.32; N, 5.57; N, 9.86. Found: C,50.38; H, 5.12; N, 9.96.

Example 16.54-Amino-7-(cis-5′-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.45 (10% MeOH in CH₂Cl₂). mp 135-138° C. Anal Calcd forC₂₁H₂₄ClN₄O₇P.0.2H₂O.0.4 CH₂Cl₂: C, 46.87; H, 4.63; N, 10.22. Found: C,47.02; H, 4.25; N, 9.99.

Example 16.64-Amino-7-(cis-5′-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineMethanesulfonic Acid Salt

R_(f)=0.45 (10% MeOH in CH₂Cl₂). mp 125-128° C. Anal Calcd forC₂₁H₂₄N₄O₇ClP.1.6 CH₃SO₃H.1.0H₂O: C, 39.76; H, 4.78; N, 8.21; S, 7.52.Found: C, 39.39; H, 4.30; N, 8.30; S, 7.96.

Example 16.74-Amino-7-(cis-5′-O-[4-(S)-(pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.40 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 183-185° C. Anal Calcd forC₂₀H₂₄N₅O₇P1.6H₂O: C, 47.45; H, 5.42; N, 13.83. Found: C, 47.78; H,5.47; N, 13.77.

Example 16.84-amino-7-(cis-5′-O-[4-(3-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.15 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₁H₂₄FN₄O₇P.0.3 H₂O: C,50.46; H, 4.96; N, 11.21. Found: C, 50.63; H, 5.35; N, 10.94.

Example 16.94-Amino-7-(cis-5′-O-[4-(3-bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

Rf=0.48 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₁H₂₄BrN₄O₇P.0.5H₂O: C, 44.70; H, 4.47; N, 9.93. Found: C, 44.58; H, 4.52; N, 9.56.

Example 16.104-Amino-7-(czs-5′-O-[4-(2-bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.15 (10% MeOH in CH₂Cl₂). mp 132-135° C. Anal Calcd forC₂₁H₂₄BrN₄O₇P.0.5H₂O: C, 44.7; H, 4.47; N, 9.93. Found: C, 44.73; H,4.69; N, 9.82.

Example 16.114-Amino-7-(cis-5′-O-[4-(5-bromopyridin-3-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.35 (10% MeOH in EtOAc) mp 132-135° C. Anal Calcd forC₂₀H₂₃N₅O₇BrP. 0.5H₂O. 0.5 EtOAc: C, 43.36; H, 4.63; N, 11.49. Found: C,43.37; H, 4.80; N, 11.16.

Example 16.124-Amino-7-(cis-5′-O-[4-(S)-phenyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.42 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 115-118° C. Anal Calcd forC₂₁H₂₅N₄O₇P. 0.4 EtOAc. 1.0H₂O: C, 51.25; H, 5.75; N, 10.58. Found: C,51.07; H, 5.88; N, 10.35.

Example 16.134-Amino-7-(cis-5′-O-[4,5-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.45 (10% MeOH in CH₂Cl₂). mp 174-177° C. Anal Calcd forC₂₉H₃₀F₃N₄O₉P.1.75H₂O: C, 49.90; H, 4.48; N, 8.03. Found: C, 49.68; H,4.82; N, 8.1.

Example 16.144-Amino-7-(cis-5′-O-[4-(2-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.48 (10% MeOH in CH₂Cl₂). mp 187-190° C. Anal Calcd forC₂₁H₂₄ClN₄O₇P. H₂O. 0.2 DMF: C, 47.72; H, 5.05; N, 10.77. Found: C,47.66; H, 5.02; N, 10.96.

Example 16.154-Amino-7-(cis-5′-O-[4-(2-fluoro-5-bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.48 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₁H₂₃BrFN₄O₇P.1.3H₂O: C,42.27; H, 4.32; N, 9.39. Found: C, 42.26; H, 4.03; N, 9.36.

Example 16.164-Amino-7-(cis-5′-O-[4,6-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.20 (10% MeOH in CH₂Cl₂). mp 140-143° C. Anal Calcd forC₂₇H₂₉N₄O₇P.1.25H₂O.CF₃CO₂H: C, 50.55; H, 4.75; N, 8.13. Found: C,50.25; H, 4.88; N, 7.99.

Example 16.174-Amino-7-(cis-5′-O-[4(3,5-bis-trifluoromethylphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.15 (10% MeOH in CH₂Cl₂). mp 130-134° C. Anal Calcd forC₂₃H₂₃N₄O₇P. 0.6H₂O: C, 44.33; H, 3.91; N, 8.99. Found: C, 44.29; H,4.13; N, 8.98.

Example 16.184-Amino-7-(trans-5′-O-[4,6-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.48 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp>220° C. Anal Calcd forC₂₇H₂₉N₄O₇P.0.9H₂O: C, 57.02; H, 5.46; N, 9.85. Found: C, 57.55; H,5.97; N, 9.88.

Example 16.194-Amino-7-(cis-5′-O-[4-(3-bromo-pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofilranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f) 0.3 (10% MeOH in EtOAc). mp 116-120° C. Anal Calcd forC₂₀H₂₃N₅O₇BrP.1H₂O. 0.6 EtOAc: C, 42.90; H, 4.79; N, 11.17 Found: C,42.90; H, 4.42; N, 10.82.

Example 16.204-Amino-7-(cis-5′-O-[4-(2,4-dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.15 (10% MeOH in CH₂Cl₂). mp 184-188° C. Anal Calcd forC₂₂H₂₄F₃N₄O₇P. 0.6H₂O: C, 47.59; H, 4.57; N, 10.09. Found: C, 47.46; H,4.96; N, 10.10.

Example 16.214-Amino-7-(cis-5′-O-[4-(3-trifluoromethylphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f) 0.15 (10% MeOH in CH₂Cl₂). mp 120-124° C. Anal Calcd forC₂₁H₂₃Cl₂N₄O₇P.0.5H₂O: C, 45.50; H, 4.36; N, 10.11. Found: C, 45.32; H,4.58; N, 10.26.

Example 16.224-Amino-7-(trans-5′-O-[4,5-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.75 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 160-163° C. Anal Calcd forC₂₇H₂₉N₄O₇P.1.2H₂O: C, 56.48; H, 5.51; N, 9.76. Found: C, 56.34, H,5.75; N, 9.71.

Example 16.234-Amino-7-(cis-5′-O-[cis-(5-methoxy-4-phenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.25 (10% MeOH in CH₂Cl₂). mp 116-120° C. Anal Calcd forC₂₂H₂₆N₄O₈PC1.1.75H₂O.1.5 CF₃CO₂H: C, 40.39; H, 4.20; N, 7.54. Found: C,39.95; H, 3.85; N, 7.38.

Example 16.244-Amino-7-(cis-5′-O-[trans-(5-methoxy-4-phenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.30 (10% MeOH in CH₂Cl₂). mp 140-143° C. Anal Calcd forC₂₂H₂(N₄O₈PC_(1.2.5)H₂O.2.2 CF₃CO₂H: C, 37.89; H, 4.00; N, 6.70. Found:C, 37.73; H, 3.61; N, 6.85.

Example 16.254-Amino-7-(cis-5′-O-[4-(2-bromo-5-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.3 (10% MeOH in CH₂Cl₂). mp 193-196° C. Anal Calcd forC₂₁H₂₃N₄O₇PClBr.1.75H₂O.1 CF₃CO₂H: C, 37.57; H, 3.77; N, 7.62. Found: C,37.20; H, 3.49; N, 7.36.

Example 16.264-Amino-7-(cis-5′-O-[4-(3,5-dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.3 (10% MeOH in CH₂Cl₂). mp 182-185° C. Anal Calcd forC₂₁H₂₃N₄O₇Cl₂P.0.3 MeOH.0.5H₂O: C, 45.37; H, 4.50; N, 9.93. Found: C,45.36; H, 4.18; N, 9.58.

Example 16.274-Amino-7-(cis-5′-O-[4-(3,5-difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.35 (10% MeOH in CH₂Cl₂). mp 135-140° C. Anal Calcd forC₂₁H₂₃N₄O₇F₂P.1.0H₂O: C, 47.55; H, 4.75; N, 10.56. Found: C, 47.29; H,4.51; N, 10.28.

Example 16.284-Amino-7-(cis-5′-O-[4-(R)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

Rf=0.45 (10% MeOH in CH₂Cl₂). mp 126-128° C. Anal Calcd forC₂₁H₂₄ClN₄O₇P.1.0H₂O: C, 47.69; H, 4.96; N, 1059. Found: C, 47.31; H,4.77; N, 10.3.

Example 16.294-Amino-7-(cis-5′-O-[4-(2-trifluoromethylphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

Rf=0.5 (10% MeOH in CH₂Cl₂). Mp 115-120° C. Anal Calcd forC₂₂H₂₄F₃N₄O₇P.1.0H₂O.1.0 CF₃CO₂H: C, 42.61; H, 4.02; N, 8.28. Found: C,42.78; H, 4.07; N, 8.27.

Example 16.304-Amino-7-(cis-5′-O-[4-(R)-(pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.3 (20% MeOH in EtOAc). mp 132-136° C. Anal Calcd forC₂₀H₂₄N₅O₇P.0.03H₂O.0.7 CH₂Cl₂: C, 46.52; H, 4.79; N, 13.14. Found: C,46.13; H, 4.39; N, 13.50.

Example 16.314-Amino-7-(cis-5′-O-[4-(3-bromo-4-fluoro-phenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.35 (10% MeOH in EtOAc). mp 122-125° C. Anal Calcd forC₂₁H₂₃N₄O₇FBrP.0.2 CF₃CO₂H: C, 43.12; H, 3.92; N, 9.40. Found: C, 42.82;H, 3.76; N, 9.57.

Example 16.324-Amino-7-(cis-5′-O-[4-(4-pyridin-3-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.30 (10% MeOH in EtOAc). mp 134-138° C. Anal Calcd forC₂₀H₂₄N₅O₇P.1.5H₂O: C, 47.62; H, 5.40; N, 13.88. Found: C, 47.89; H,5.08; N, 13.97.

Example 16.334-Amino-7-(cis-5′-O-[4-(pyridin-2-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.50 (10% MeOH in CH₂Cl₂). nip 88-90° C. Anal Calcd forC₂₀H₂₄N₅O₇P.2.3H₂O.1.3 CF₃CO₂H: C, 40.69; H, 4.52; N, 10.50. Found: C,40.38; H, 4.86; N, 10.90.

Example 16.344-Amino-7-(cis-5′-O-[4-(R)-(phenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.30 (10% MeOH in CH₂Cl₂). mp 177-180° C. Anal Calcd forC₂₁H₂₅N₄O₇P. 0.1 EtOAc. 0.2 CF₃CO₂H: C, 51.54; H, 5.16; N, 11.03. Found:C, 51.92; H, 4.78; N, 10.75.

Example 16.354-Amino-7-(cis-5′-O-[4-(4-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofanosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.45 (10% MeOH in CH₂Cl₂). mp 182-184° C. Anal Calcd forC₂₁H₂₄N₄O₇ClP.2.0H₂O.2.9 CF₃CO₂H: C, 36.68; H, 3.55; N, 6.38. Found: C,36.33; H, 3.35; N, 6.44.

Example 16.364-Amino-7-(cis-5′-O-[4-(2,3-difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.5 (10% MeOH in CH₂Cl₂). mp 177-180° C. Anal Calcd forC₂₁H₂₃F₂N₄O₇P.1.9H₂O.1.1CF₃CO₂H: C, 41.46; H, 4.18; N, 8.34. Found: C,42.07; H, 4.02; N, 8.68.

Example 16.374-Amino-7-(cis-5′-O-[4-(2-fluoro-5-methoxyphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.4 (10% MeOH in CH₂Cl₂). mp 80-85° C. Anal Calcd forC₂₂H₂₆N₄O₈FP.0.4H₂O.2.0 CF₃CO₂H: C, 41.11; H, 3.82; N, 7.37. Found: C,41.13; H, 3.50; N, 7.54.

Example 16.384-Amino-7-(cis-5′-O-[4-(2-chloro-4-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

Rf=0.46 (15% MeOH in CH₂Cl₂). mp 138-141° C. Anal Calcd forC₂₁H₂₃ClFN₄O₇P. 0.3H₂O. 0.9 CF₃CO₂H: C, 43.00; H, 3.88; N, 8.80. Found:C, 42.73; H, 4.21; N, 8.55.

Example 16.394-Amino-7-(cis-5′-O-[4-(2-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

Rf=0.48 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 101-103° C. Anal Calcd forC₂₁H₂₄FN₄O₇P. 1.5H₂O: C, 48.37; H, 5.22; N, 10.74. Found: C, 48.70; H,5.47; N, 10.43.

Example 16.404-Amino-7-(cis-5′-O-[4-(2-cyanophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

Rf=0.42 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₂H₂₄N₅O₇P. 2H₂O.0.1 CF₃CO₂H: C, 48.58; H, 5.16; N, 12.76. Found: C, 48.86; H, 5.51; N,12.70.

Example 16.414-Amino-7-(cis-5′-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

R_(f)=0.45 (10% MeOH in CH₂Cl₂). mp 145-148° C. Anal Calcd forC₂₁H₂₄N₄O₇PC1.0.7 CH₂Cl₂.1.2 CF₃CO₂H: C, 40.93; H, 3.79; N, 7.92; F,9.67. Found: C, 40.43; H, 3.77; N, 8.22; F, 9.47.

Example 16.424-Amino-7-(cis-5′-O-[4-phenyl-5,6-tetramethylene-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.24 (15% MeOH in,CH₂Cl₂-1% NH₄OH). mp 110-113° C. Anal Calcd forC₂₄H₃₁N₄O₇P. 2.0H₂O: C, 53.00; H, 6.23; N, 9.89. Found: C, 53.03; H,5.93; N, 9.91.

Example 16.434-Amino-7-(cis-5′-O-[4-(3-cyanophenyl)-2-oxo-1,3,2-dioxaphosphornan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyriridine

R_(f)=0.51 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 157-160° C. Anal Calcd forC₂₂H₂₄NsO₇P. 2.5H₂O: C, 48.35; H, 5.35; N, 12.82. Found: C, 48.50; H,5.72; N, 12.77.

Example 16.444-Amino-7-(cis-5′-O-[4-(3,4-dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.25(10% MeOH in CH₂Cl₂). Anal Calcd for C₂₁H₂₃N₄O₇PCl₂.0.2H₂O.0.3 EtOAc: C, 46.34; N, 4.52; N, 9.74. Found: C, 46.00; H, 4.26; N,9.43.

Example 16.454-Amino-7-(cis-5′-O-[4-(S)-(3-pyridyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.3 (10% MeOH in EtOAc).

Example 16.464-Amino-7-(cis-5′-O-[4-(S)-(3-pyridyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.3 (10% MeOH in EtOAc).

Example 16.474-Amino-7-(cis-5′-O-[4-phenyl-2-oxo-6-spirocyclohexyl-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.35 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₆H₃₃N₄O₇P.0.2H₂O.0.2CF₃CO₂H.0.2 EtOAc: C, 55.51; H, 6.03; N, 9.52. Found: C, 55.72; H, 5.87;N, 9.18.

Example 16.484-Amino-7-(cis-5′-O-[4-phenyl-2-oxo-6-spirocyclopentyl-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.6 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₅H₃₁N₄O₇P.1.0 F₃CO₂H:C, 50.31; H, 5.00; N, 8.69. Found: C, 49.99; H, 4.99; N, 8.68.

Example 16.494-Amino-7-(cis-5′-O-[4,4-Dimethyl-6-(4-pyridyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.35 (15% MeOH in CH₂Cl₂-1% NH₄OH). MH⁺ Calcd for C₂₂H₂₈N₅O₇P:506. Found: 506.

Example 16.504-Amino-7-(cis-5′-O-[4-(4-cyanophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.40 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₂H₂₄N₅O₇P.2.2H₂O: C, 48.67; H, 5.31; N, 12.90. Found: C, 48.74; H,5.61; N, 12.54.

Example 16.514-Amino-7-(cis-5′-O-[6-(3-chlorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.58 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₃H₂₈ClN₄O₇P.2.0H₂O: C, 48.05; H, 5.61; N, 9.74. Found: C, 48.36; H, 5.74; N, 9.62.

Example 16.524-Amino-7-(cis-5′-O-[4-(3-chlorophenyl)-2-oxo-6-spirocyclopropyl-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.35 (12% MeOH in CH₂Cl₂). Anal Calcd for C₂₅H₂₇ClF₃N₄O₉P. 1.6H₂O.0.5 CH₂Cl₂: C, 42.26; H, 4.34; N, 7.72. Found: C, 41.95; H, 3.95; N,7.43.

Example 17 Preparation of Prodrugs of 2′-C-beta-methyl-7-deazaguanosinevia trans-phosphate Addition

The parent nucleoside2-amino-7-(2-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-onewas synthesized as described in US2002-0147160A1 and WO 02/057827.

The nucleoside was converted to corresponding prodrug following theprocedures as in steps A, B and C of Example 16.

The following examples were synthesized as described steps A-C.

Example 17.12-Amino-7-(cis-5′-O-[4-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphospborinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.30 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₁H₂₄ClN₄O₈P.1.2CF₃CO₂NH₄.1.0 CF₃CO₂H: C, 38.22; H, 3.76; N, 9.13. Found: C, 37.93; N,3.80; N, 9.40.

Example 17.22-Amino-7-(cis-5′-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-Cmethyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f) 0.15 (10% MeOH in CH₂Cl₂). mp 175° C. Anal Calcd forC₂₁H₂₄ClN₄O₈P.0.5H₂O: C, 47.07; H, 4.70; N, 10.46. Found: C, 46.73; H,4.90; N, 10.16.

Example 17.32-Amino-7-(cis-5′-O-[4-(5-bromo-2-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-Cmethyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin4(3H)-one

R_(f)=0.41 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₁H₂₃BrFN₄O₈P.0.5H₂O. 0.2 CF₃CO₂H: C, 41.38; H, 3.93; N, 9.02. Found: C, 41.60; H,4.32; N, 8.77.

Example 17.42-Amino-7-(cis-5′-O-[4-(3-bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-Cmethyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-oneTrifluoroacetic Acid Salt

R_(f)=0.38 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 142-145° C. Anal Calcd forC₂₁H₂₄N₄O₈P. 0.7H₂O. 0.9 CF₃CO₂H: C, 39.89; H, 3.86; N, 8.16. Found: C,39.53; H, 3.65; N, 8.43.

Example 17.52-Amino-7-(cis-5′-O-[4-(3-Chloro-4-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrTolo[2,3-d]pyrimidin-4(3H)-one

R_(f) 0.45 (20% MeOH in CH₂Cl₂. Anal Calcd for C₂₁H₂₃N₄O₈FCIP. 1.4H₂O:C, 44.24, H, 4.78; N, 9.83. Found: C, 43.77; H, 4.78; N, 10.31.

Example 17.62-Amino-7-(cis-5′-O-[4-(2,5-difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.35 (20% MeOH in CH₂Cl₂). mp 170-173° C. Anal Calcd forC₂₁H₂₃F₂N₄O₈P.2.0H₂O.0.4 CF₃CO₂NH₄: C, 42.45; H, 4.67; N, 9.99. Found:C, 42.28; H, 4.76 N, 9.96.

17.7:2-Amino-7-(cis-5′-O-[4-(2-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.25 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₁H₂₄ClN₄OSP.1.25H₂O. 0.2 CF₃CO₂H: C, 44.92; H, 4.70; N, 9.79. Found: C, 44.93; H,5.09; N, 10.08.

17.8:2-Amino-7-(cis-5′-O-[4-(pyridin-2-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-Cmethyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-oneTrifluoroacetic Acid Salt

R_(f)=0.4 (15% MeOH in CH₂Cl₂). mp 180-190° C. Anal Calcd forC₂₀H₂₄N₅O₈P.1.3 CF₃CO₂H.0.3 CH₂Cl₂: C, 41.23; H, 3.91; N, 10.50. Found:C, 40.96; H, 3.46; N, 11.05.

17.9:2-Amino-7-(cis-5′-O-[4-(2-trifluoromethylphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-Cmethyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-oneTrifluoroacetic Acid Salt

R_(f)=0.4 (10% MeOH in CH₂Cl₂). mp 185-188° C. Anal Calcd forC₂₂H₂₄N₄O₈F₃P.0.8 CF₃CO₂H: C, 43.50; H, 3.84; N, 8.60. Found: C, 43.55;H, 3.97; N, 8.98.

17.10:2-Amino-7-(cis-5′-O-[4-(R)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-oneTrifluoroacetic Acid Salt

Rf=0.50 (15% MeOH in CH₂Cl₂). mp 170-180° C.

Example 17.112-Amino-7-(cis-5′-O-[4-(3,5-difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-oneTrifluorbacetic Acid Salt

R_(f)=0.30 (10% MeOH in CH₂Cl₂) mp 182-185° C. Anal Calcd forC₂₁H₂₃N₄OSF₂P.0.3 EtOAc. 0.2 CF₃CO₂H: C, 46.99; H, 4.47; N, 9.70. Found:C, 47.26; H, 4.32; N, 9.46.

Example 17.122-Amino-7-(cis-5′-O-[4-(3,5-dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f) 0.35 (10% MeOH in CH₂Cl₂). mp 177-180° C. Anal Calcd forC₂₁H₂₃N₄O₈C₁₂P.0.1 EtOAc.0.2 CF₃CO₂H. C, 44.16; H. 4.08; N, 9.45. Found:C, 44.33; H, 4.44; N, 9.18.

Example 17.132-Amino-7-(cis-5′-O-[4-(S)-(pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.21 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 138-141° C., Anal Calcd forC₂₀H₂₄N₅O₈P. 2.2H₂O: C, 45.07; H, 5.33; N, 13.14. Found: C, 45.12; H,5.40; N, 12.89.

Example 17.142-Amino-7-(cis-5′-O-[4-(3-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin4(3H)-one

R_(f)=0.25 (10% MeOH in CH₂Cl₂). mp 170° C. Anal Calcd forC₂₁H₂₄FN₄O₈P.1.5H₂O: C, 46.93; H, 5.06; N,10.42. Found: C, 46.92; H,5.12; N, 10.44.

Example 17.152-Amino-7-(cis-5′-O—[4-(3-bromo-4-fluoro-phenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.25 (10% MeOH in CH₂Cl₂). mp 175-179° C. Anal Calcd forC₂₁H₂₃BrFN₄O₈P. 0.5,H₂O. 0.5 EtOAc: C, 43.01; H, 4.39; N, 8.72. Found:C, 43.03; H, 4.49; N, 8.49.

Example 17.162-Amino-7-(cis-5′-O-[4-(R)-phenyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.30 (10% MeOH in CH₂Cl₂) mp 128-133° C. Anal Calcd forC₂₁H₂₅N₄O₉P. 1.1H₂O.0.3 CF₃CO₂H: C, 47.48; H, 5.07; N, 10.25. Found: C,47.61; H, 5.36; N, 9.91.

Example 17.172-Amino-7-(cis-5′-O-[4,5-cis-diphenyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-oneTrifluoroacetic Acid Salt

R_(f)=0.45 (20% MeOH in CH₂Cl₂). mp 187-190° C. Anal Calcd forC₂₇H₂₉N₄O₈P.2H₂O.1.3 CF₃CO₂H: C, 47.23; H, 4.59; N, 7.44. Found: C,46.83; H, 4.33; N, 7.31.

Example 17.182-Amino-7-(cis-5′-O-[6,6-dimethyl-4-phenyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-oneTrifluoroacetic Acid Salt

R_(f)=0.40 (20% MeOH in CH₂Cl₂). mp 192-194° C. Anal Calcd forC₂₃H₂₉N₄O₈P.2.0H₂O.1.0 CF₃CO₂H: C, 44.78; H, 5.11; N, 8.36. Found: C,44.40; H, 4.67; N, 8.22.

Example 17.192-Amino-7-(cis-5′-O-[cis-(5-methoxy-4-phenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.30 (20% MeOH in CH₂Cl₂). mp 148-151° C. Anal Calcd forC₂₂H₂₆N₄O₉ClP.1.0H₂O: C, 45.96; H, 4.91; N, 9.75. Found: C, 46.03; H,4.80; N, 9.64.

Example 17.202-Amino-7-(cis-5′-O-[4-(2,3-difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-oneTrifluoroacetic Acid Salt

R_(f)=0.5 (10% MeOH in CH₂Cl₂). mp 215-220° C. Anal Calcd forC₂₁H₂₃N₄O₈F₂P.1.0H₂O.1.0 CF₃CO₂H: C, 41.83; H, 3.97; N, 8.48. Found: C,41.70; H, 3.77; N, 8.50.

Example 17.212-Amino-7-(cis-5′-O-[4-(2-bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.15 (10% MeOH in CH₂Cl₂). mp 180° C. Anal Calcd forC₂₁H₂₄BrN₄O₈P.1.1H₂O: C, 42.67; H, 4.47; N, 9.48. Found: C, 42.51, H,4.60; N, 9.58.

Example 17.222-Amino-7-(cis-5′-O-[4-(3,4-dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f) 0.30 (10% MeOH in CH₂Cl₂). Mp 192-195° C. Anal Calcd forC₂₁H₂₃N₄OSCl₂P.0.2 CF₃CO₂H. 0.2 EtOAc: C, 44.31; H, 4.15; N, 9.31.Found: C, 44.40; H, 3.94; N, 9.21.

Example 17.232-Amino-7-(cis-5′-O-[4-(3,5-bis-(trifluoromethylphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.15 (10% MeOH in CH₂Cl₂) Mp 155-175° C. Anal Calcd forC₂₃H₂₃F₆N₄O₈P.0.6H₂O: C, 43.22; H, 3.82; N, 8.76. Found: C, 43.08; H,4.03; N, 8.94.

Example 17.242-Amino-7-(cis-5′-O-[4-(3-trifluoromethylphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.15 (10% MeOH in CH₂Cl₂). mp 145-165° C. Anal Calcd forC₂₂H₂₄F₃N₄O₈P.1H₂O: C, 45.68; H, 4.53; N, 9.69. Found: C, 45.31; H,4.88; N, 9.71.

Example 17.252-Amino-7-(cis-5′-O-[4-(2,4-dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.15 (10% MeOH in CH₂Cl₂) mp 175° C. Anal Calcd forC₂₁H₂₃Cl₂N₄O₈P.1H₂O: C, 43.54; H, 4.35; N, 9.67. Found: C, 43.32; H,4.35; N, 9.55.

Example 17.262-Amino-7-(cis-5′-O-[4-(5-bromo-pyridin-3-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.3 (10% MeOH in CH₂Cl₂) mp 185-189° C. Anal Calcd forC₂₀H₂₃N₅O₈BrP.1.5 CF₃CO₂H: C, 37.16; H, 3.32; N, 9.42. Found: C, 37.23;H, 3.44; N, 9.33.

Example 17.272-Amino-7-(cis-5′-O-[4(yridin-3-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidin-4(3H)-one

R_(f)=0.15 (10% MeOH in CH₂Cl₂); Anal Calcd for C₂₀H₂₄N₅Os₈P.1 H₂O.0.4EtOAc: C, 47.46; H, 5.38; N, 12.81. Found: C, 47.40; H, 5.17; N, 12.78.

Example 185′-O-[4-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyladenosine

2′-C-methyl adenosine was made as described in WO 01/90121.

Step A:

General procedure for synthesis of cyclic phosphoramidites fromsubstituted diols:

To a solution of commercially available diisopropyl phosphoramidousdichloride (1 nmol) in THF (5 mL) was added 1,3-diol (1 mmol) andtriethylamine (4 mmol) in THF (5 mL) at −78° C. over 30 min. Thereaction was slowly warmed to room temperature and left stirringovernight. Reaction mixture was filtered to remove salts and filtratewas concentrated to give crude product. Silica gel column chromatographyprovided pure cyclic diisopropyl phosphoramidite of 1,3-diol.

Step B:

General procedure for nucleoside-cyclic phosphoramidite coupling andoxidation (J. Org. Chem. 61:7996 (1996)):

To a solution of nucleoside (1 mmol) and cyclic phosphoramidite (1 mmol)in DMF (10 mL) was added benzimidazolium triflate (1 mmol). The reactionwas stirred for 30 min at room temperature. The mixture was cooled to−40° C. before addition of t-butylhydro peroxide (2 mmol) and left atroom temperature overnight. Concentration under reduced pressure andchromatography of crude product resulted in pure cyclic propyl prodrug.

R_(f)=0.46 (12% MeOH in CH₂Cl₂). mp 153° C. Anal calcd forC₂₀H₂₃ClN₅O₇P: C, 46.93; H, 4.53; N, 13.63. Found: C, 47.06; H, 4.36; N,13.68.

Example 19 cis-5′-O-[4-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan2-yl]-2′-C-methyl-guanosine

2′-C-Methyl guanosine was made as described in WO 01/90121.

The nucleoside was converted to corresponding prodrug following theprocedures as in steps A, B and C of Example 16.

R_(f)=0.35 (25% MeOH in CH₂Cl₂). mp>230° C. Anal calcd forC₂₀H₂₃ClN₅O₈P: C, 45.51; H, 4.39; N, 13.27. Found: C, 45.89; H, 4.44; N,13.23.

Example 20cis-5′-O-[4-(S)-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-guanosine

The compound was synthesized in a similar sequence as Example 19 usingthe phosphorylating agent whose preparation is described in Example 14.

R_(f)=0.35 (20% MeOH in CH₂Cl₂). mp>180° C. Anal calcd forC₂₀H₂₃N₅O₈ClP.1.0H₂O.0.8 CF₃CO₂H: C,40.72; H, 4.08; N, 10.99. Found: C,40.43; N, 4.41; N, 11.34.

Example 21 Preparation of prodrugs of 2′-C-beta-methyl-adenosine viatrans-phosphate Addition

2′-C-methyl adenosine was made as described in WO 01/90121.

The nucleoside was converted to corresponding prodrug following theprocedures as in steps A, B and C of Example 16.

trans-phosphorylating agents utilized in step B are synthesized by theprocedures as described in examples 1-15.

Example 21.1cis-5′-O-[4-(S)-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-adenosineTrifluoroacetic Acid Salt

R_(f)=0.3 (5% MeOH in EtOAc). mp 125-128° C. Anal calcd forC₂₀H₂₃ClN₅O₇P.1.7 CF₃CO₂H: C, 39.83; H, 3.53; N, 9.92. Found: C, 39.52;H, 3.46; N, 10.21.

Example 21.2cis-5′-O-[4-(3-Cyanophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.43 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 153-156° C. Anal calcd forC₂₁H₂₃N₆O₇P.1.1H₂O: C, 48.30; H, 4.86; N, 16.09. Found: C, 48.53; H,5.11; N, 15.75.

Example 21.3cis-5′-O-[4-(2,5-Difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-adenosine

R_(f) 0.60 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 75-78° C. Anal calcd forC₂₀H₂₂F₂N₅O₇P.0.3 CH₂Cl₂: C, 45.25; H, 4.23; N, 13.00. Found: C, 45.07;H, 3.94; N, 12.69.

Example 21.4cis-5′-O-[4-(3,5-Difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.65 (15% MeOH in CH₂Cl₂-1% NH₄OH). mp 120-123° C. Anal calcd forC₂₀H₂₂F₂N₅O₇P.1.5H₂O.0.1 C₆H₁₄: C, 45.07; H, 4.85; N, 12.76. Found: C,45.04; H, 5.25; N, 12.59.

Example 21.5cis-5′-O-[4-(S)-(Pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.55 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal calcd forC₁₉H₂₃N₆O₇P.2.5H₂O: C, 43.60; H, 5.39; N, 16.06. Found: C, 43.35; H,5.54; N, 16.05.

Example 21.6cis-5′-O-[4-(3-Bromophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyladenosine

R_(f)=0.5 (10% MeOH in CH₂Cl₂). mp 108-110° C. Anal calcd forC₂GH₂₃N₅O₇BrP.1.5H₂O.0.4 CF₃CO₂H: C, 39.72; H, 4.23; N, 11.14. Found: C,39.44; H, 4.55; N, 11.18.

Example 21.7cis-5′-O-[4-(Pyridin-2-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-adenosineTrifluoroacetic Acid Salt

R_(f)=0.4 (10% MeOH in CH₂Cl₂). mp 118-120° C. Anal calcd forC₁₉H₂₃N₆O₇P.2.0H₂O.1.0 CF₃CO₂H: C, 40.14; H, 4.49; N, 13.37. Found: C,40.36; H, 4.92; N, 13.63.

Example 21.8cis-5′-O-[4-(4-Methylsulfonylphenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-adenosineTrifluoroacetic Acid Salt

R_(f)=0.3 (10% MeOH in CH₂Cl₂). mp 185-187° C. Anal calcd forC₂₁H₂₆N₅O₉PS.0.6H₂O.0.6 CF₃CO₂H: C, 42.01; H, 4.41; N, 11.03. Found: C,41.93; H, 4.73; N, 10.97.

Example 21.9cis-5′-O-[4-(Pyridine-3-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.2 (10% MeOH in EtOAc). mp.137-140° C. Anal calcd forC₁₉H₂₃N₆O₇P.1.5H₂O.0.4 EtOAc. C, 45.76; H, 15.54; N, 5.44. Found: C,45.88; H, 15.19; N, 5.09.

Example 21.10cis-5′-O-[4-(5-Bromo-3-pyridyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.15 (10% MeOH in EtOAc). Anal Calcd for C₁₉H₂₂N₆O₇BrP.1.0H₂O.0.4EtOAc: C, 40.52; H, 4.49; N, 13.76. Found: C, 40.39; H, 4.22; N, 13.42.

Example 21.11cis-5′-O-[4-(2-Bromophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.35 (5% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₃BrN₅O₇P.1.5 H₂O.0.1CH₂Cl₂: C, 40.79; H, 4.46; N, 11.83. Found: C, 40.49; H. 4.46; N, 11.49.

Example 21.12cis-5′-O-[4-(3-Methylsulfonylphenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]2-C-beta-methyl-adenosine

R_(f)=0.3 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₁H₂₆N₅O₉PS.1.4 H2O.1.0CH₂Cl₂: C, 39.70; H, 4.66; N, 10.52. Found: C, 39.61; H, 4.11; N, 10.22.

Example 21.13cis-5′-O-[4-(3,5-Dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-beta-C-methyl-adenosineTrifluoroacetic Acid Salt

R_(f)=0.15 (10% MeOH in EtOAc). Anal Calcd for C₂₀H₂₂N₅O₇Cl₂P.1.0H₂O.1.0 CF₃CO₂H: C, 38.95; H, 3.71; N, 10.32. Found: C, 38.56; H, 3.52;N, 10.57.

Example 21.14cis-5′-O-[4-(3-Fluorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-y]-2′-C-beta-methyl-adenosine

R_(f)=0.5 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₃N₅O₇FP.0.4 CF₃CO₂H:C, 46.18; H, 4.36; N, 12.94. Found: C, 46.09; H, 4.39; N, 13.01.

Example 21.15cis-5′-O-[6-(3-Chlorophenyl)-4,4-dimethyl-2-oxo-1,3-dioxaphosphosphorin-2-yl]-2′-beta-methyl-adenosine

R_(f) 0.50 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₂H₂₇ClN₅O₇P.1.0H₂O.0.5 CH₃OH: C, 47.09; H. 5.44; N, 12.20. Found: C,47.00; H, 5.81; N, 12.21.

Example 21.16cis-5′-O-[4-(3,4-Dichloro)-2-oxo-1,3,2-dioxaphosphorin-2-y]-2′-C-beta-methyl-adenosineTrifluoroacetic Acid Salt

R_(f) 0.40 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₂N₅O₇Cl₂P.1.7CF₃CO₂H. 2.7H₂O: C, 35.63; H, 3.72; N, 8.88. Found: C, 35.17; H, 3.55;N, 8.80.

Example 21.17cis-5′-O-[4-(3-Fluoro,4-chloro)-2-oxo-1,3,2-dioxaphosphorin-2-y]-2′-C-beta-methyl-adenosine

R_(f) 0.45 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₂N₅O₇PCIF. 0.8CF₃CO₂H.0.9H₂O: C, 40.71; H, 3.89; N, 10.99. Found: C, 40.46; H, 3.93;N, 10.98.

Example 21.18cis-5′-O-[4-(3-Acetophenyl)-2-oxo-1,3,2-dioxaphosphosphorin-2-yl]-2′-beta-methyl-ladenosine

R_(f)=0.40 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₂H₂₆N₅O₈P.0.4CH₂Cl₂.1.0H₂O: C, 47.08; H, 5.08; N, 12.26. Found: C, 47.03; H, 4.94; N,12.15.

Example 21.19cis-5′-O-{4-[3-(Morpholine-4-sulfonyl)phenyl]-2-oxo-1,3,2-dioxaphosphosphorin-2-yl}-2′-beta-methyl-adenosine

R_(f)=0.60 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₄H₃₁N₆O₁₀PS.0.6H₂O. 0.5 CH₂Cl₂: C, 43.28; H, 4.92; N, 12.36. Found: C,43.65; H, 4.88; N, 11.98.

Example 21.20cis-5′-O-{4,4-Dimethyl-6-(4-pyridyl)-2-oxo-1,3,2-dioxaphosphosphorin-2-yl}-2′-beta-methyl-adenosine

R_(f)=0.40 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₁H₂₇N₆O₇P.0.9H₂O. 0.4 CH₂Cl₂: C, 46.18; H, 5.36; N, 15.10. Found: C,46.00; H, 4.98; N, 15.09.

Example 21.21cis-5′-O-[4-(R)-(3-Chlorophenyl)-1,3,2-dioxaphosphoran-2-yl]-2′-beta-methyl-adenosine

R_(f)=0.3 (5% MeOH in EtOAc). Anal Calcd for C₂₀H₂₃N₅O₇ClP.1.0 H₂O.0.2EtOAc: C, 45.63; H, 4.90; N, 12.79. Found: C, 45.53; H, 4.75; N, 12.50.

Example 21.22cis-5′-O-[4-(2,3-Difluorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-y]-2′-C-beta-methyl-adenosine

R_(f)=0.35 (15% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₂N₅O₇F₂P.0.75H₂O:C, 45.59; H, 4.50; N, 13.29. Found: C, 45.49; H, 4.08; N, 13.30.

Example 21.23cis-5′-O-[4-(R)-(4-Pyridyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-beta-C-methyl-adenosine

R_(f)=0.3 (20% MeOH in EtOAc). Anal Calcd for C₁₉H₂₃N₆O₇.1.7H₂O: C,44.83; H, 5.23; N, 16.51. Found: C, 44.73; H, 5.06; N, 16.30.

Example 21.24cis-5′-O-[4,4-Dimethyl-6-phenyl-2-oxo-1,3,2-dioxaphosphoran-2-yl]-2′-beta-methyl-adenosineTrifluoroacetic Acid Salt

R_(f)=0.3(10% MeOH in EtOAc). Anal Calcd for C₂₂H₂gN₅O₇P.1.0 H₂O.1.5CF₃CO₂H. 0.1 EtOAc: C, 43.38; H, 4.63; N, 9.96. Found: C, 43.38; H,4.71; N, 9.71.

Example 21.25cis-5′-O-[4-(4-Cyanophenyl)-2-oxo-1,3,2-dioxaphosphosphorin-2-yl]-2′-beta-methyl-adenosine

R_(f)=0.60 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₁H₂₃N₆O₇P.1.0H2O.0.1 EtOAc: C, 48.47; H, 4.84; N, 16.15. Found: C,48.89; H, 4.42; N, 15.68.

Example 21.26cis-5′-O-[4-Phenyl-2-oxo-6-spirocyclohexyl-1,3,2-dioxaphosphorin-2-yl]-2′-beta-C-methyl-adenosineTrifluoroacetic Acid Salt

R_(f)=0.3 (10% MeOH in EtOAc). Anal Calcd for C₂₁H₂₃N₆O₇P.1.0 H₂O.0.1EtOAc: C, 44.69; H, 5.02; N, 9.31. Found: C, 44.40; H, 5.00; N, 9.39.

Example 21.27cis-5′-O-{4-(4-Fluoro-3-trifluoromethylphenyl)-2-oxo-1,3,2-dioxaphosphosphorin-2-yl}-2′-beta-methyl-adenosine

R_(f)=0.55 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₁H₂₂F₄N₅O₇P.H₂O: C, 43.38; H, 4.16; N, 12.05. Found: C, 43.41; H, 3.85; N, 12.04.

Example 21.28cis-5′-O-{4-[3-(2-Furanyl)pyridyl]-2-oxo-1,3,2-dioxaphosphorin-2-yl}-2′-C-beta-methyl-adenosine

R_(f)=0.5 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₃H₂₅N₆O₈P.3.0 H₂O.0.1CF₃CO₂H: C, 45.69; H, 5.14; N, 13.78. Found: C, 45.64; H, 5.03; N,14.05.

Example 21.29cis-5′-O-{4-[3-(2-Thiophenyl)pyridyl]-2-oxo-1,3,2-dioxaphosphorin-2-yl}-2′-C-beta-methyl-adenosine

R_(f)=0.55 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₃H₂₅N₆O₇SP.2.3 H₂O.2.0CF₃CO₂H: C, 39.07; H, 3.84; N, 10.13. Found: C, 38.70; H, 3.64; N,10.28.

Example 21.30cis-5′-O-[4-(2-Methoxy-pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.3 (15% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₅N₆O₈P.1.0CF₃CO₂H.1.2H₂O: C, 41.03; H, 4.44; N, 13.05. Found: C, 40.96; H, 4.97;N, 13.70.

Example 21.31cis-5′-O-[4,4-Dimethyl-6-(3,4-dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.25 (10% MeOH in CH₂Cl₂). Anal Calcd forC₂₂H₂₆Cl₂N₅O₇P.1.3H₂O.0.6 CH₂Cl₂: C, 41.70; H, 4.62; N, 10.74. Pound: C,41.57; H, 4.78; N, 10.83.

Example 21.32cis-5′-O-[6-(3,5-Difluorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.28 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₂H₂₆F₂N₅O₇P.2.0H₂O.0.3 CF₃CO₂H: C, 44.38; H, 4.99; N, 11.45. Found: C, 44.56; H, 5.18;N, 11.21.

Example 21.33cis-5′-O-[4-(2-Bromo-5-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-beta-C-methyl-adenosineTrifluoroacetic Acid Salt

R_(f)=0.3 (10% MeOH in EtOAc). Anal Calcd for C₂₀H₂₂N₅O₇BrFP.3.4H₂O. 2.2CF₃CO₂H. 0.1 EtOAc: C, 33.27; H, 3.58; N, 7.82. Found: C, 32.94; H,3.24; N, 7.52.

Example 21.34cis-5′-O-[4,4-Dimethyl-6-(3-fluorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-beta-methyl-adenosine

R_(f)=0.5 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₂H₂₇N₅O₇FP.1.8 H₂O.1.5CF₃CO₂H: C, 41.31; H, 4.45; N, 9.63. Found: C, 40.94; H, 4.50; N, 9.38.

Example 21.35cis-5′-O-[4,4-Dimethyl-6-(2,3-difluorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-y]-2′-C-beta-methyl-adenosine

R_(f)=0.4 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₂H₂₆F₂N₅O₇P.0.5 H₂O: C,48.00; H, 4.94; N, 12.72. Found: C, 47.62; H, 4.90; N, 12.67.

Example 21.36cis-5′-O-[6,6-Dimethyl-4-(3,5-dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-beta-C-methyl-adenosineTrifluoroacetic Acid Salt

R_(f)=0.2 (10% MeOH in EtOAc). MH⁺ Calcd for C₂₂H₂₆Cl₂N₅O₇P: 575. Found:575.

Example 21.37cis-5′-O-{4-[4-(2-Furanyl)pyridyl]-2-oxo-1,3,2-dioxaphosphorin-2-yl}-2′-C-beta-methyl-adenosine

R_(f)=0.5 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₃H₂₅N₆O₈P.1.5 H₂O.1.5CF₃CO₂H: C, 42.06; H, 4.00; N. 11.32. Found: C, 41.67; H, 4.30; N,11.13.

Example 21.38cis-5′-O-{[4-(2-thiomethyl-pyrdin-4-yl)-2-oxo-1,3,2-dioxaphos-phorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.3 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₂H₂₆F₃N₆O₉PS.2.4 H₂O:C, 38.76; H, 4.55; N, 12.33. Found: C, 38.39; H, 4.12; N, 12.09.

Example 21.39cis-5′-O-[4-(2-cyanopyridin-3-yl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.3 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₂N₇O₇P.0.5H₂O. 2.2CF₃CO₂H: C, 38.40; H, 3.33; N, 12.85. Found: C, 38.09; H, 3.25; N,12.57.

Example 21.40cis-5′-O-[4,4-Diethyl-6-phenyl-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.55 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₄H₃₂N₅O₇P.0.3H₂O: C, 53.49; H, 6.10; N, 13.00. Found: C, 53.97; H,6.40; N, 12.61.

Example 21.41cis-5′-O-[4-(5-Methyl-3-pyridyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-adenosine

R_(f)=0.2 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₅N₆O₇P.1.2H₂O: C,46.73; H, 5.37; N, 16.35. Found: C, 46.64; H, 5.21; N, 16.15.

Example 21.42cis-5′-O-[6-(5-Bromo-2,3-difluorophenyl)-4,4-dimethyl-1-2-oxo-1,3,2-dioxa-phosphorin-2-yl]-2′-C-methyl-adenosine

R_(f)=0.45 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₂H₂₅BrF₂N₅O₇P.1.0 CH₃OH: C, 42.34; H, 4.48; N, 10.73. Found: C, 42.82;H, 4.84; N, 10.66.

Example 23 General Procedure for Preparation of 2′,3′-cyclic CarbonateProdrugs of 2′-C-beta-methyl-7-deazaadenosine Prodrugs

To a solution of 5′-substituted cyclic propyl prodrug (0.25 mmol) in DME(2.5 mL)was added carbonyl diimidazole (CDI) (0.5 mmol) at 0° C. Thereaction was warmed to room temperature and stirred for 4 h. Solvent wasremoved under reduced pressure and the crude product was chromatographedto give 2′,3′-carbonate as a solid.

Example 23.14-Amino-7-(2′,3′-carbonyl-cis-5′-O-[4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.45 (10% MeOH in CH₂Cl₂). mp 127-130° C. Anal calcd forC₂₂H₂₂N₄O₈PCl.1.0H₂O: C, 47.62; H, 4.36; N, 10.10. Found: C, 47.94; H,4.10; N, 10.13.

Example 23.24-Amino-7-(2′,3′-carbonyl-cis-5′-O-[4-(S)-(pyridin-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidine

R_(f)=0.4 (20% MeOH in CH₂Cl₂). mp 192-195° C. Anal calcd forC₂₁H₂₂N₅O₈P.1.0 H₂O: C, 48.37; H, 4.64; N, 13.43. Found: C, 48.41; H,4.39; N, 13.60.

Example 26

Parent nucleoside is prepared as described in US2002-0147160A1, WO02/057827.

Prodrugs are synthesized as described in steps A, B and C of example 16.Phosphorylating agents were made as described in examples 11-16.

Example 26.1cis-5′-O-[4-(S)-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-beta-C-methyl-2-amino-7-deaza-adenosineTrifluoroacetic Acid Salt

R_(f)=0.3 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₁H₂₅N₅O₇ClP.2.0CF₃CO₂H: C, 39.83; H, 3.61; N, 9.29. Found: C, 39.70; H, 3.57; N, 9.55.

Example 26.2cis-5′-O-[4-(3,5-Dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-beta-C-methyl-2-amino-7-deaza-adenosine

R_(f)=0.2 (10% MeOH in EtOAc). Anal Calcd for C₂₁H₂₄Cl₂N₅O₇P.1.0H₂O.0.25 CH₃OH: C, 43.53; H, 4.64; N, 11.94. Found: C, 43.50; H, 4.25;N, 11.55.

Example 26.3cis-5′-O-[4-(3-Pyridyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-beta-C-methyl-2-amino-7-deaza-adenosine

R_(f)=0.2 (15% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₅N₆O₇P.1.2 H₂O: C,46.73; H, 5.37; N, 16.35. Found: C, 46.41; H, 5.02; N, 16.14.

Example 26.4cis-5′-O-[6-(-2,3-Difluorophenyl)-4,4-dimethyl-1,2,3-dioxa-2-oxo-phosphorin-2-yl]-2′-C-methyl-2-amino-7-deaza-adenosine

R_(f)=0.40 (150/oMeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd for C₂₃H₂₈F₂N₅O₇P.0.8H₂O: C, 48.47; H, 5.24; N, 12.29. Found: C, 48.67; H, 5.39; N, 11.94.

Example 26.5cis-5′-O-[4-(2,3-Difluorophenyl)-1,2,3-dioxa-2-oxo-phosphorinan-2-yl]-2′-C-methyl-2-amino-7-deaza-adenosine

R_(f)=0.60 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₁H₂₄F₂N₅O₇P.1.0H₂O: C, 46.24; H, 4.80; N, 12.84. Found: C, 46.12; H,4.87; N, 12.63.

Example 26.6cis-5′-O-(4,4-Dimethyl-6-phenyl-1,2,3-dioxa-2-oxo-phosphorin-2-yl)-2′-C-methyl-2-amino-7-deaza-adenosine

R_(f)=0.64 (15% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₃H₃₀N₅O₇P.0.8H₂O: C, 51.74; H, 5.97; N, 13.12. Found: C, 51.91; H,5.90; N, 12.75.

Example 26.7cis-5′-[4-(4-(S)-Pyridyl)-1,2,3-dioxa-2-oxo-phosphorin-2-yl]-2′-C-methyl-2-amino-7-deaza-adenosine

R_(f)=0.3 (20% MeOH in CH₂Cl₂-1% NH₄OH). Anal Calcd forC₂₀H₂₅N₆O₇P.1.3H₂O.1.1 CF₃CO₂H: C, 41.58; H, 4.51; N, 13.11. Found: C,41.14; H, 4.10; N, 13.59.

Example 27 Example 27.12,4-Diamino-5-fluoro-7-beta-D-(5-(4-(S)-(3-Chlorophenyl)-1,3-dioxa-2-oxophosphorinan-2-yl)-2-methylribofuranosyl)pyrrolo[2,3-d]pyrimidine

Prodrug was prepared as described in steps A and B of example 18.

MH⁺ Calcd for C₂₁H₂₄ClFN₅O₇P: 544. Found: 544. Example 28

The prodrug was synthesized as described in Example 21.

General procedure for N⁶-carbamate formation (Bioorg. Med. Chem. 8:1697(2000)):

To a solution of prodrug (1 mmol) in dry THF (6 mL) and pyridine (4 mL)was added n-pentyl chloroformate (0.37 mL, 2 mmol) dropwise at 0° C.over a period of 15 min. The mixture was warmed to room temperature andstirred for an additional 30 min before quenching the reaction withmethanol (3 mL). The reaction mixture was then concentrated underreduced pressure and the product was purified by silica gel columnchromatography.

Example 28.1cis-5′-O-[4-(S)-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-beta-methyl-N-6-n-pentylcarbamoyl-adenosinetrifluoroacetic acid

R_(f)=0.6 (5% MeOH in CH₂Cl₂). Anal Calcd for C₂₆H₃₃N₅O₉ClP.0.8 CF₃CO₂H:C, 46.22; H, 4.75; N, 9.76. Found: C, 46.00; H, 4.84; N, 9.97.

Example 30

5′-monophosphate prodrugs were made as described in Example 21.2′,3′-carbonates were prepared following the procedure described inExample 23.

Example 30.1cis-5′-O-[4-(S)-Pyrid-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.4 (15% MeOH in CH₂Cl₂). Anal calcd for C₂₀H₂₁N₆O₈P.1.2H₂O: C,45.67; H, 4.48; N, 15.98. Found: C, 45.18; H, 3.99; N, 15.74.

Example 30.2cis-5′-O-[4-(S)-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.45 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₁H₂₁N₅O₈ClP.1.0 H₂O: C,45.38; H, 4.17; N, 12.60. Found: C, 45.21; H, 3.97; N, 12.41.

Example 30.3cis-5′-O-[4-(3-Fluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.6 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₁H₂₁N₅O₈FP.0.7 CH₂Cl₂: C,44.87; H, 3.89; N, 12.06. Found: C, 44.67; H, 3.86; N, 12.01.

Example 30.4cis-5′-O-[4-(2,3-Difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-y]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.35 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₁H₂₀N₅O₈F₂P.1.0 H₂O: C,45.25; H, 3.98; N, 12.56. Found: C, 44.88; H, 3.74; N, 12.47.

Example 30.5cis-5′-O-[6-(3-Chlorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.42 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₃H₂₅ClN₅O₈P.1.0 H₂O: C,47.31; H, 4.66; N, 11.99. Found: C, 47.15; H, 4.83; N, 11.95.

Example 30.6cis-5′-O-[6-Phenyl-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf 0.50 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₃H₂₆N₅O₈P. 0.5 H₂O: C,51.11; H, 5.04; N, 12.96. Found: C, 51.16; H, 5.28; N, 13.09.

Example 30.7cis-5′-O-[4-(3,4-Dichlorophenyl)-2-oxo-1,3,2-dioxaphosphoran-2-yl]-2′,3′-carbonyl2′-C-methyl-adenosine

Rf=0.35 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₁H₂₀C₁₂N₅O₈P: C, 44.07;H, 3.52; N, 12.39. Found: C, 43.68; H, 3.90; N, 12.43.

Example 30.8cis-5′-O-[4-(3,5-Difluorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.40 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₁H₂₀F₂N₅O₈P.1.5H₂O: C,44.53; H, 4.09; N, 12.36. Found: C, 44.31; H, 3.75; N, 12.18.

Example 30.9cis-5′-O-[6-(3,5-Difluorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.39 (10% MeOH in CH₂Cl₂). Ana calcd for C₂₃H₂₄F₂N₅O₈P. 0.3 CH₂Cl₂:C, 47.20; H, 4.18; N, 11.81. Found: C, 47.56; H, 3.84; N, 11.51.

Example 30.10cis-5′-O-[6-(2,3-Difluorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-y]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.35 (5% MeOH in CH₂Cl₂). Anal calcd for C₂₃H₂₄F₂N₅O₈P. 0.6H₂O: C,47.77; H, 4.39; N, 12.11. Found: C, 47.30; H, 3.92; N, 11.90.

Example 30.11cis-5′-O-[6-(3,4-Dichlorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.45 (10% MeOH in CH₂Cl₂). MH+ Calcd for C₂₃H₂₄Cl₂N₅OgP: 601. Found:601.

Example 30.12cis-5′-O-[6-(3-Fluorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.4 (5% MeOH in CH₂Cl₂). Anal calcd for C₂₃H₂₅N₅O₈P.0.4 H₂O: C,49.63; H, 4.67; N, 12.58. Found: C, 49.43; H, 4.60; N, 12.71.

Example 30.13cis-5′-O-[6-yrid-4-yl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.32 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₂H₂₅N₆O₈P.2.0H₂O: C,46.48; H, 5.14; N, 14.78. Found: C, 46.30; H, 4.80; N, 14.56.

Example 30.14cis-5′-O-[4-(3,5-Dichlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.3 (10% MeOH in EtOAc). Anal calcd for C₂₁H₂₀ClN₅O₈P.1.0H₂O.0.5Imidazole: C, 43.28; H, 3.87; N, 13.46. Found: C, 43.28; H, 3.92; N,13.79.

Example 30.15cis-5′-O-[6-(3,5-Dichlorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf 0.3 (5% MeOH in EtOAc). Anal calcd for C₂₃H₂₄Cl₂N₅O₈P: C, 46.02; H,4.03; N, 11.67. Found: C, 45.39; H, 3.10; N, 10.79.

Example 30.16cis-5′-O-[6-(5-Bromo-2,3-difluorophenyl)-4,4-dimethyl-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.5 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₃H₂₃BrF₂N₅O₈P.1.2 CH₂Cl₂:C, 38.85; H, 3.42; N, 9.36. Found: C, 38.51; H, 3.38; N, 9.66.

Example 30.17cis-5′-O-[4-(5-Bromo-pyrid-4-yl)-2-oxo-1,3,2-dioxaphosphor-an-2-yl]-2′,3′-carbonyl-2′-C-methyl-adenosineTrifluoroacetic Acid Salt

Rf=0.30 (10% MeOH in CH₂Cl₂). Anal Calcd for C₂₀H₂₀N₆O₈BrP.0.9

H₂O.1.0 CF₃CO₂H: C, 37.03; H, 3.22; N, 11.78. Found: C, 36.68; H, 3.11;N, 12.15.

Example 31 General Procedure for Preparation of 2′,3′-carbonateNucleosides via 5′-protected Nucleosides Step A:

To a solution of nucleoside analog (0.5 mmol) in DMF (5 mL) was addedimidazole (1.5 mmol) followed by t-butyldimethylsilyl chloride (0.6mmol) at 0° C. The reaction was allowed to warm to room temperature andstirred for 3 h. The mixture was evaporated under reduced pressure. Theresidue was extracted with CH₂Cl₂, washed with water and dried. Theorganic extract was evaporated and the product was purified by columnchromatography.

Step B:

To a solution of 5′-t-butyldimethylsilyloxy-protected nucleoside (0.25mmol) in DMF (2.5 mL) was added carbonyl diimidazole (CDI) (0.5 mmol) at0° C. The reaction was warmed to room temperature and stirred for 3 h.The solvent was removed under reduced pressure and the crude product waschromatographed to give the 2′,3′-carbonate as a solid.

Step C:

The 5′-t-butyldimethylsilyloxy-protected nucleoside 2′,3′-carbonate(0.15 mmol) was dissolved in a pre-cooled 75% aq.TFA (3 mL) and allowedto stir at 0° C. for 2 h. The reaction mixture was evaporated underreduced pressure. The crude product was purified by flashchromatography.

Example 31.1 8-bromo-2′,3′-carbonyl-2′-C-methyl-adenosine

Rf=0.65 (10% MeOH in CH₂Cl₂). Anal calcd for C₁₂H₁₂N₅O₅Br.0.2 CH₃OH: C,37.33; H, 3.29; N, 17.84. Found: C, 3.7.48; H, 3.37; N, 17.45.

Example 31.24-Amino-7-(2′,3′-carbonyl-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo[2,3-d]pyrimidineTrifluoroacetic Acid Salt

Rf=0.5 (10% MeOH in CH₂Cl₂). Anal calcd for C₁₃H₁₄N₄O₅. 1.0 H₂O.2.0CF₃CO₂H: C, 36.97; H, 3.28; N, 10.14. Found: C, 37.18; H, 3.10; N, 9.80.

Example 31.3 2′,3′-Carbonyl-2′-C-methyl-cytidine Trifluoroacetic AcidSalt

Rf=0.2 (10% MeOH in CH₂Cl₂). Anal calcd for C₁₁H₁₃N₃O₆. 0.8H₂O.0.9CF₃CO₂H: C, 38.41; H, 3.90; N, 10.50. Found: C, 38.14; H, 3.72; N,10.77.

Example 31.4 2′,3′-Carbonyl-2′-C-beta-methyl-inosine

R_(f)=0.25 in 20% MeOH-dichloromethane. Anal Calcd for C₁₂H₁₂N₄O₆.0.3CF₃CO₂H.0.1C₂H₅O: C, 45.02; H, 3.88; N, 16.28. Found: C, 44.63; H, 3.65;N, 16.15

Example 31.5 2′,3′-Carbonyl-2′-C-beta-methyl-adenosine

Rf=0.5 (10% MeOH in CH₂Cl₂). Anal calcd for C₁₂H₁₃N₅O₅. 0.7 CF₃CO₂H: C,41.58; H, 3.57; N, 18.09. Found: C, 41.26; H, 3.42; N, 18.02.

Example 31.6 2′,3′-Carbonyl-2′-beta-C-methylguanosine

Rf=0.1 (10% MeOH in CH₂Cl₂). Rf=0.25 (10% MeOH in CH₂Cl₂). Anal Calcdfor C₁₂H₁₃N₅O₆. 0.2 CF₃CO₂H: C, 43.04; H, 3.84; N, 20.24. Found: C,43.15; H, 3.86; N, 20.52.

Example 31.7 2′,3′-Carbonyl-4′-C-methyl-cytidine

Rf=0.45 (15% MeOH in CH₂Cl₂). Anal calcd for C₁₁H₁₃N₃O₆. 0.8 CF₃CO₂H: C,40.42; H, 3.71; N, 11.22. Found: C, 40.26; H, 3.77; N, 11.60.

Example 32 General Procedure for Preparation of 2′,3′-carbonateNucleosides via Single-Pot 5′-protection and 2′,3′-carbonylation Step A:

To a solution of a nucleoside analog (0.5 mmol) in DMF (5 mL) was addedimidazole (1.5 mmol) followed by t-butyldimethylsilyl chloride (0.6mmol) at 0° C. The reaction was allowed to warm to room temperature andstirred for 3 h. Carbonyl diimidazole (CDI) (0.6 mmol) was added to thereaction at 0° C. upon consumption of all the starting material. Thereaction was then warmed to room temperature and stirred for anadditional 3 h. The reaction was evaporated under reduced pressure. Themixture was extracted with CH₂Cl₂, washed with water and dried. Theorganic extract was evaporated and the product was purified bychromatography.

Step B:

Same as Step C of example 31.

Example 32.14,6-Diamino-7-(2′,3′-Carbonyl-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo-[2,3-d]pyrimidine

Rf=0.4 (10% MeOH in CH₂Cl₂). Anal calcd for C₁₃H₁₅N₅O₅. 0.6 CF₃CO₂H: C,43.77; H, 4.03; N, 17.99. Found: C, 43.51; H, 3.97; N, 17.60.

Example 33

Procedure for single-step synthesis of 2′,3′-carbonate nucleosides.

Example 31.5 was also made by the following procedure.

To a solution of 2′-C-methyl-adenosine (28 mg, 0.1 mmol) in DMF (2 mL)was added diphenyl carbonate (32 mg, 0.15 mmol). The reaction was heatedto 250° C. in a sealed tube under microwave conditions for 5 min. Themixture was concentrated under reduced pressure and chromatographed byelution with 5 to 10% MeOH in CH₂Cl₂ to obtain 13 mg of desired product.

Example 34 General Procedure for Preparation of NMP Prodrugs from2′,3′-carbonate Substituted Nucleosides

To a solution of 2′,3′-carbonate nucleoside (0.25 mmol) in DMF (1.5 mL)was added t-butyl magnesium chloride and the reaction mixture wasstirred under nitrogen for 30 min. The reaction mixture was then cooledto 55° C. and the phosphorylating agent (prepared as described inexamples 14b and 15a) (0.35 mmol) in DMF (1.5 mL) was added dropwise.The reaction was allowed to warm to room temperature and stirred undernitrogen for 2 h. The reaction mixture was evaporated under reducedpressure and quenched with saturated aq.NH₄Cl solution. The mixture wasextracted with 10% MeOH in CH₂Cl₂, washed with water and dried. Theorganic extract was evaporated and the product was purified bychromatography.

Example 34.14,6-Diamino-7-[(cis-5′-O-4-(S)-(3-chlorophenyl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl)-2′,3′-carbonyl-2′-C-methyl-beta-D-ribofuranosy]-7H-pyrrolo-[2,3-d]pyrimidineTrifluoroacetic Acid Salt

Rf=0.6 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₂H₂₃N₅O₈ClP. 1.1 CF₃CO₂H:C, 42.92; H, 3.59; N, 10.34. Found: C, 42.49; H, 3.37; N, 10.23.

Example 34.24,6-Diamino-7-(cis-5′-O-[4-(S)-(pyrid-4-yl)-2-oxo-1,3,2-dioxaphosphorinan-2-yl]-2′,3′-carbonyl-2′-C-methyl-beta-D-ribofuranosyl)-7H-pyrrolo-[2,3-d]pyrimidineTrifluoroacetic Acid Salt

Rf=0.4 (15% MeOH in CH₂Cl₂). Anal calcd for C₂₁H₂₃N₆O₈P. 2.3 CF₃CO₂H: C,39.39; H, 3.27; N, 10.77. Found: C, 39.97; H, 2.96; N, 10.70.

Example 35

2′-C-Methyl-cytidine was made as described in WO 04/052899.

Step A:

2′-C-Methyl-cytidine was converted to corresponding 2′,3′-acetonidefollowing the procedure as in step A of Example 16.

Step B:

General procedure for dimethylaminomethylene protection of amine:

To a solution of 2′,3′-acetonide of 2′-C-methyl-cytidine (1.4 g, 4.71mmol) in pyridine (30 mL) was added N,N-dimethylformamide dimethylacetal (0.8 mL, 5.87 mmol). The reaction was stirred at room temperatureovernight. The mixture was then concentrated under reduced pressure. Thecrude product was chromatographed on a silica gel column eluting with 5%MeOH in dichloromethane to obtain 900 mg of dimethylamino-methyleneadduct.

Step C:

Prodrug formation was carried-out utilizing the procedure as in step Bof Example 16.

trans-phosphorylating agents utilized in step C were synthesized by theprocedures as described in examples 14 and 15.

Step D:

The amine protected 4-pyridyl prodrug (0.15 g) obtained from the abovestep was dissolved in pre-cooled 75% TFA/H₂O (10 mL) and allowed to stirat 0° C. for 8 h. The reaction mixture was evaporated under reducedpressure. The crude product was purified by flash chromatography (1%aq.NH₄OH in 10% MeOH in CH₂Cl₂) to give 0.1 g of the deprotected prodrugas an off-white solid.

Example 35.1cis-5′-O-[4-(S)-(Pyrid-4-yl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-methyl-cytidineTrifluoroacetic Acid Salt

R_(f) 0.2 (10% MeOH in CH₂Cl₂). MH⁺ Calcd for C1H₂₃N₄O₈P: 455. Found:455.

Example 35.2cis-5′-O-[4-(S)-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′-C-methyl-cytidineTrifluoroacetic Acid Salt

R_(f)=0.3 (10% MeOH in CH₂Cl₂). Anal calcd for C₁₉H₂₃N₃O₈ClP.0.5H₂O. 0.3CF₃CO₂H: C, 44.33; H, 4.61; N, 7.91. Found: C, 44.39; H, 4.42; N, 7.84.

Example 36

2′,3′-Carbonylation of Example 35.1 was performed following theprocedure described as in Example 23.

Example 36.1cis-5′-O-[4-(S)-(Pyrid-4-yl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′,3′-carbonyl-2′-C-methyl-cytidineTrifluoroacetic Acid Salt

R_(f)=0.3 (10% MeOH in CH₂Cl₂). Anal calcd for C₁₉H₂₁N₄O₉P.1.7H₂O. 2.0CF₃CO₂H: C, 37.38; H, 3.60; N, 7.58. Found: C, 37.17; H, 3.23; N, 7.97.

Example 36.2cis-5′-O-[4-(S)-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphorin-2-yl]-2′,3′-carbonyl-2′-C-methyl-cytidineTrifluoroacetic Acid Salt

R_(f)=0.35 (10% MeOH in CH₂Cl₂). Anal calcd for C₂₀H₂₁N₃O₉P.1.6Imidazole.1.4H₂O. 0.2 CF₃CO₂H: C, 45.23; H, 4.34; N, 12.98. Found: C,45.11; H, 4.12; N, 13.31.

Example 38

4′-C-Methyl-cytidine was made as described in WO 01/90121.

2′,3′-Carbonate of 4′-C-methyl-cytidine was synthesized as described inexample 31 and 5′-monophosphate prodrug was prepared as in the case ofExample 34.

Example 38.1cis-5′-O-[4-(S)-(4-Pyridyl)-2-oxo-1,3,2-dioxaphosphoran-2-yl]-2′,3′-carbonyl-4′-C-alpha-methylcytidine

R_(f) 0.45 in 40% MeOH-acetone. MH⁺ Calcd for C₁₉H₂₁N₄O₉P: 481. Found:481.

Example 38.2cis-5′-O-[4-(S)-(3-Chlorophenyl)-2-oxo-1,3,2-dioxaphosphoran-2-yl]-2′,3′-carbonyl-4′-C-alpha-methylcytidine

Rf=0.3 in 20% MeOH-dichloromethane. MH⁺ Calcd for C₂₀H₂₁N₃O₉ClP: 514.Found: 514. Anal calcd for C₂₀H₂₁N₃O₉PCl. 1.0 CF₃CO₂H. 0.2H₂O: C, 42.11;H, 3.69; N, 6.58. Found: C, 41.89; H, 3.63; N, 6.77.

Biological Examples

Examples of use of the compositions and methods of the invention includethe following. It will be understood that these examples are exemplaryand that the method of the invention is not limited solely to theseexamples.

For the purposes of clarity and brevity, chemical compounds are referredto as synthetic example numbers in the biological examples below.

Example A Conversion of Compound 31.5 to 2′-β-methyladenosine in LiverS9 from Various Species

The rate of Compound 31.5 conversion to nucleoside was evaluated inliver S9 obtained from rat, dog, monkey and human liver.

Methods:

Rat liver S9 was prepared from a freshly harvested liver homogenized inice-cold 100 mM potassium phosphate buffer, pH 7.4. The homogenate wascentrifuged in a Sorvall RC-5B Refrigerated Superspeed Centrifuge (30minutes at 10,000 g) and the resulting supernatant used in the assays.The other S9 preparations were purchased from 1n Vitro Technologies.

Compound 31.5 was incubated in 100 mM potassium phosphate buffer, pH 7.4containing liver S9 (1-10 mg/mL protein) for up to 120 minutes in anEppendorf Thermomixer R (37° C., 850 rpm). Aliquots of the reactionmixture were quenched with 2 volumes of ice-cold acetonitrile/0.2%formic acid (pH 3.0) at appropriate time intervals, briefly vortexed,and then immediately placed on dry ice. After all time points weretaken, the samples were centrifuged in an Eppendorf microfuge (14,000rpm, 10 minutes). The supernatant fractions were analyzed fordisappearance of prodrug (and appearance of 2′-β-methyladenosine) byreverse phase HPLC on an Agilent Zorbax SB-Aq column (511m, 4.6×150 mm).The loading mobile phase buffer (Buffer A) consisted of a 9:1 ratio(v/v) of 20 mM potassium phosphate, pH 6.2 and acetonitrile. The columnwas eluted with a gradient from Buffer A to 60% acetonitrile over 16minutes at a 1.0 mL/min flow rate. The elution of reaction componentswas monitored at 260 mm. Quantitation was by comparison to authenticstandards prepared in liver S9 and processed in an identical fashion tothe unknown samples. The rate of prodrug conversion was calculated fromthe linear portion of the reaction curves and expressed as nmolesconverted per mg of S9 protein per minute of reaction time.

Results:

The rate of Compound 31.5 activation in liver S9 from the variousspecies tested is shown in the table below:

Specific activity*, S9 Source nmoles/min/mg Rat liver 0.28 ± 0.03 Beagleliver 2.0 ± 0.3 Monkey liver 0.024 ± 0.003 Human liver  0.03 ± 0.005*rate of prodrug disappearance

Conclusions:

The S9 fraction from rat, dog, monkey and human liver catalyzed theconversion of Compound 31.5 to 2′-β-methyladenosine. Liver from all fourspecies thus expresses the enzymatic machinery required for carbonateprodrug conversion.

Example B HepDirect-carbonate Prodrug Activation in Rat LiverMicrosomes. Byproduct Capture Assay

Prodrugs were tested for activation in rat or human liver microsomes bymeans of a prodrug byproduct capture assay.

Methods:

Prodrugs were tested for activation by rat or human liver microsomespurchased from In Vitro Technologies. The study was performed at 2 mg/mLliver microsomes, 100 mM KH₂PO₄, 10 mM glutathione, 25 μM or 250 μMcompound, and 2 mM NADPH for 0-7.5 min. in an Eppendorf Thermomixer 5436(37° C., 850 rpm). The reactions were initiated by addition of NADPHfollowing a 2-min. preincubation. Reactions were quenched with 60%methanol at 0, 2.5, 5, and 7.5 min.L-Glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine, aglutathione adduct of the by-product of prodrug activation,3-chlorophenyl vinyl ketone, was quantified following extraction of thereaction with 1.5 volumes of methanol. The extracted samples werecentrifuged at 14,000 rpm in an Eppendorf microfuge and the supernatantanalyzed by HPLC forL-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycinecontent. SpikedL-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycinestandards (1-30 μM) were prepared in 2 mg/mL microsomes under reactionconditions and then quenched and processed in an identical fashion tounknown samples. For HPLC analysis, the loading mobile phase buffer(Buffer A) consisted of a 9:1 ratio (v/v) of 20 mM potassium phosphate,pH 6.2 and acetonitrile. Extract (100 μL) was injected onto a BeckmanUltrasphere ODS column (4.6×250 mM). The column was eluted with agradient to 60% acetonitrile. The elution ofL-glutamyl-L-(S-(3-oxo-3-(3-chlorophenyl)propyl)cysteinylglycine(retention time 10.4 min.) was monitored at 245 nm.

Results:

The rate of enzymatic cleavage of the HepDirect prodrug moiety ofCompound 34.1 as assessed by the generation of the glutathione captureproduct assay in rat and human liver microsomes (HLM and RLM,respectively) is shown in the table below:

HLM activation rate, RLM activation rate, 34.1 concentrationnmoles/min/mg nmoles/min/mg  25 μM 0.68 ± 0.02 0.13 ± 0.01 250 μM 0.59 ±0.1   0.22 ± 0.024

Conclusions:

Rat and human liver microsomes catalyze the removal of the HepDirectprotecting group from HepDirect-carbonate prodrugs.

Example C Kinetics of Conversion of HepDirect-Carbonate Prodrugs toNucleoside Monophosphates by Human Liver Microsomes

The kinetics of activation of prodrug analogues to NMP were evaluated inthe microsomal fraction of human liver.

Methods:

Human liver microsomes were purchased from In Vitro Technologies. Thestudy was performed at 2 mg/mL human liver microsomes, 100 mM KH₂PO₄, 10mM glutathione, 0-250 μM compound, and 2 mM NADPH for 0-7.5 min. in anEppendorf Thermomixer 5436 (37° C., 850 rpm). The reactions wereinitiated by addition of NADPH following a 2-min. preincubation.Reactions were quenched with 60% methanol at 0, 2.5, 5, and 7.5 min. Theresulting extracts were clarified by centrifugation at 14,000 rpm in anEppendorf microfuge (20 min.). The supernatants (200 μL) were evaporatedunder vacuum and, heat to dryness. The dried residue was reconstitutedwith 200 μL of water and the mixture was centrifuged for 10 min at14,000 rpm. A mixture of 35 μL aliquot of supernatant and 35 μL ofmobile phase A (20 mM N—N-dimethylhexylamine and 10 mM propionic acid in20% methanol) was analyzed by LC-MS/MS (Applied Biosystems, API 4000)equipped with an Agilent 1100 binary pump and a LEAP injector. NMP wasdetected by using MS/MS mode (M⁻/78.8) and quantified based oncomparison to a standard of lamivudine monophosphate. Kinetic parameterswere determined from a Lineweaver-Burke plot of the data with use ofSigmaPlot 9.0 software.

Results:

Compound Km, μM Vmax, nmoles/min/mg 30.1 64 0.36 30.10 44.5 0.12

Conclusions:

The HepDirect-carbonate prodrugs evaluated (Compounds 30.1 and 30.10)were activated to the corresponding NMP in human liver microsomes,indicating that the enzymes required for removal of both the HepDirectand the carbonate prodrug moieties are present in this reaction system.In addition, the affinity of the prodrugs for the enzyme systems washigh, as indicated by the relatively low apparent Km values.

Example D HepDirect-carbonate Prodrug Activation in Human LiverMicrosomes. LC-MS Assay

The activation of prodrug analogues to NMP was evaluated in themicrosomal fraction of human liver.

Methods:

NMP generation was evaluated in human liver microsomes essentially asdescribed above in Example C. A single concentration of each prodrug wastested (25 μM).

Results:

The rates of activation of the prodrugs evaluated, expressed as nmolesof NMP generated per minute per milligram of microsomal protein, areshown in the table below:

Compound nmoles/min/mg 30.1 0.2 30.2 0.23 30.3 0.13 30.4 0.11 30.5 0.0130.6 0.01 30.7 0.04 30.8 0.1 30.9 0.02 30.10 0.03 30.11 0 30.12 0.0130.13 0.02 30.14 0.01 30.15 0.01

Conclusions:

The prodrugs evaluated were generally well activated to generate the NMPin human liver microsomes.

Example E Susceptibility of an 2′-β-C-Methyladenosine and its2′,3′-Carbonate and HepDirect Prodrugs to Non-Productive Metabolism InVitro and In Vivo

The susceptibility of 2′-β-C-methyladenosine, and its 2′,3′-carbonateprodrugs to non-productive metabolism was evaluated in vitro and invivo.

Methods:

In Vitro—2′-β-C-methyladenosine and Compounds 31.5, 30.1, and 30.10 wereseparately incubated in heparinized whole rat blood or plasma at 37° C.Aliquots of the blood and plasma samples were removed periodically andextracted with perchloric acid and methanol, respectively, and thencentrifuged. The acidic supernatants were neutralized with potassiumcarbonate and recentrifuged. The neutralized supernatants and methanolicextracts were then analyzed for the major metabolite of2′-β-C-methyladenosine, i.e., 2′-β-C-methylinosine, as described below.

The methanolic plasma extracts were analyzed by HPLC using an Agilent1100. Analysis (50 μL) was performed on an Agilent Zorbax SB-Aq column(4.6×150 mm) eluted with a gradient consisting of a mixture of Buffer A(20 mM potassium phosphate pH 6.2) and Buffer B (acetonitrile) (0-10min, 0-10%; 10-20 min, 10-80%, 20-21 min, 80-0%; 21-30 min, 0-0%) and UVabsorbance monitoring at 265 nm. The flow rate was 1.5 mL/min and thecolumn temperature was set at 40° C. Concentrations of metabolite weredetermined from calibration curves prepared by spiking known amounts ofstandards to plasma and processing as before. The LOQ of2′-β-C-methylinosine was 1 μM.

In Vivo—2′-β-C-Methyladenosine and Compounds 31.5, 30.1, and 30.10 wereadministered by IV bolus and oral gavage to separate sets of maleSprague-Dawley rats. At pre-specified times at 20 min, 1, 3, 5, 8, 12,16, and 24 hrs post dose, liver, heart, and thigh muscle samples (˜1 g)were harvested by freeze-clamping and homogenized in 3 volumes ofice-old acetonitrile. Following centrifugation to clarify thehomogenate, aliquots of the tissue extracts were analyzed for deaminatedproducts, such as 2′-B-C-methylinosine, and other nucleoside metabolitesby HPLC as described in Example E. Also at the pre-specified times,blood (heparinized) was collected from the vena cava and centrifugedbriefly to obtain plasma. Following extraction of the plasma with 1.5volumes of methanol, the samples were vortexed and centrifuged to removethe precipitate. The supernatant was then analysis for deaminatedproducts and other metabolites by LC-UV as described above. The areaunder the curve (AUC) to the last measurable time point was calculatedby trapezoidal summation of the plasma, liver, heart, and muscleconcentration-time profile of the major metabolite 2′-β-C-methylinosine.

Results:

While 2′-β-C-methylinosine was observed after incubation of2′-β-C-methyladenosine and Compound 31.5 in rat whole blood in vitro, nodeaminated products were detected following the incubation of Compounds30.1 and 30.10. After IV or oral administration of Compound 31.5 torats, 2′-B-C-methylinosine was detected in plasma, liver, heart, andmuscle tissue but no or only trace levels of the deaminated product weremeasurable following dosing of Compounds 30.1 and 30.10 as summarized inthe table below (IV data shown only).

AUC 2′-MeIno* AUC AUC 2′-MeIno* Muscle 2′-MeIno* COMPOUND Heart nmol ·hr/g nmol · hr/g Plasma μM · hr 2′-β- nd nd 26.8 Methyladenosine 31.513.4 2.6 5.5 30.1 <2.9 <2.9 <7.2  30.10 <2.9 <2.9 <7.2 nd = notdetermined *dose normalized to 5.5 mg/kg nucleoside equivalents

Conclusions:

Prodrugs of 2′-β-C-methyladenosine, Compounds 30.1 and 30.10, weresignificantly less susceptible to deamination and other non-productivemetabolism both in vitro and in vivo when compared to the freenucleoside. Compound 31.5 also showed reduced susceptibility todeamination relative to 2′-β-C-methyladenosine. The improved enzymaticstability profiles are expected to provide enhanced bioavailability of2′-β-methyladenosine and consequently the delivery of higher levels ofthe active phosphorylated form of the nucleoside to the target organ(liver).

Example F NTP Generation in Hepatocytes Following Incubation withNucleoside Analogues and their Prodrugs

Nucleoside analogues and their prodrugs were evaluated for their abilityto generate NTPs in freshly isolated rat hepatocytes.

Methods:

Hepatocytes were prepared from fed Sprague-Dawley rats (250-300 g)according to the procedure of Berry and Friend (Berry, M. N. Friend, D.S., J. Cell Biol. 43:506-520 (1969)) as modified by Groen (Groen, A. K.et al., Eur. J. Biochem 122:87-93 (1982)). Hepatocytes (20 mg/mL wetweight, >85% trypan blue viability) were incubated at 37° C. in 2 mL ofKrebs-bicarbonate buffer containing 20 mM glucose, and 1 mg/mL BSA for 2h in the presence of 1-250 μM nucleoside or prodrug (from 25 mM stocksolutions in DMSO). Following the incubation, 1600 μL aliquot of thecell suspension was centrifuged and 300 μL of acetonitrile was added tothe pellet, vortexed and sonicated until the pellet broke down. Then 200μL of water was added to make a 60% acetonitrile solution. After 10 mincentrifugation at 14,000 rpm, the resulting supernatant was transferredto a new vial and evaporated to near dryness in a Savant SpeedVac Plusat room temperature: The dried residue was reconstituted with 200 μL ofwater and the mixture was centrifuged for 10 min at 14,000 rpm. Amixture of 35 μL aliquot of supernatant and 35 μL of mobile phase A (20mM N—N-dimethylhexylamine and 10 mM propionic acid in 20% methanol) wasanalyzed by LC-MS/MS (Applied Biosystems, API 4000) equipped with anAgilent 1100 binary pump and a LEAP injector. NTP was detected by usingMS/MS mode (M⁻/78.8) and quantified based on comparison to a standard oflamivudine triphosphate.

Results:

The amount of NTP generated over a 2 hour period in rat hepatocytesfollowing incubation of nucleosides and prodrugs at a concentration of25 μM, is shown in the table below:

NTP concentration, No. nmoles/g 23.2 130.8 30.1 185 30.2 295.5 30.3 31330.4 279.5 30.5 43.3 30.6 132.6 30.7 96.7 31.5 183 30.8 113 30.11 1.930.9 39.1 30.10 31.5 30.12 12.9 30.13 53.3 30.14 48.8 30.15 18.8 34.18.7 32.1 0.1 34.2 8.8 31.2 22.5 31.3 0 30.16 36 36.1 118.6 31.7 0 38.116.6 38.2 214.3 31.6 8.7 30.17 60.5

Conclusions:

Compounds of this invention showed an ability to generate NTP in freshlyisolated rat hepatocytes. It is generally accepted that the NTP form ofa nucleoside is the active antiviral agent.

Example G Antiviral Activity of Nucleosides and Prodrugs in HCV-InfectedHuman Liver Slice Assay

The effect of various compounds on HCV replication in human liver tissuewas evaluated as described below.

Methods:

Liver from a brain-dead HCV antibody-positive human patient was perfusedwith ice-cold Viaspan (Dupont Pharmaceutical) preservation solution andreceived on ice in Viaspan.

Precision-cut liver slices of ˜200-250 μm thickness and 8 cm diameterwere prepared and cultured in Waymouth's cell culture medium (Gibco,Inc.) that was supplemented with 10% fetal bovine serum and 10 mL/LFungi-Bact at 37° C., and gassed with carbogen (95% O₂, 5% CO₂) at 0.75liters/min. Tissue slices were maintained in culture for 72 h. Cellculture medium containing test compound in solution was changed on adaily basis.

At appropriate times of liver slice incubation, liver slices and mediumwere collected for analysis of HCV RNA (tissue and medium) andnucleotide levels (NTP). All collected media and tissue slices weremaintained in liquid N₂ until analysis.

Medium and tissue samples were analyzed for HCV RNA levels according topublished procedures (Bonacini et. al., 1999) which utilize anautomated, multicycle, polymerase chain reaction (PCR)-based technique.This assay has a lower limit of detection for HCV RNA of 100 viralcopies/ml. Frozen liver slices were disrupted by using a combination ofultrasound probe sonication, Branson Sonifier 450 (Branson Ultrasonics,Danbury, Conn.) and homogenization using a Dounce conical pestle in 200μls of 10% (v/v) perchloric acid (PCA). After a 5 min centrifugation at2,500×g, the supernatants were neutralized using 3 M KOH/3 M KHCO₃ andmixed thoroughly. The neutralized samples were centrifuged at 2,500 gfor 5 min and NTP levels were determined by ion exchange phase HPLC(Hewlett Packard 1050) using a Whatman Partisil 5 SAX (5 μm, 4.6×250 mm)column. Samples (50 μL) were injected onto the column in 70% 10 mMammonium phosphate buffer and 30% 1 M ammonium phosphate buffer, both atpH 3.5 and containing 6% ethanol. Nucleoside triphosphates were elutedfrom the column using a linear gradient to 80% 1 M ammonium phosphate pH3.5/6% ethanol buffer, at a flow rate of 1.25 mL/min and detected by UVabsorbance (254 nm).

Results:

HCV RNA levels present in the liver slice culture media decreased fromthe levels present in control, untreated slices following incubationwith 2′-β-methyladenosine and Compounds 30.1 and 31.5 (see table below).Dose-dependent formation of the corresponding NTP was observed intreated slices. Liver slice ATP levels were large unaffected bytreatment.

Concentration, 2′-β-Me- Compound Compound μM Adenosine* 30.1* 31.5* 0.250 0 0 2.5 83 0 93 25 94 94 85 100 92 96 75 *% decrease in viral RNArelative to untreated liver slices

Conclusions:

2′-β-methyladenosine and Compounds 30.1 and 31.5 reduced hepatitis Cvirus RNA levels in the HCV-infected human liver slice assay.Dose-dependent conversion of nucleoside and prodrugs to the active NTPwas observed in slices. Assuming good pharmacokinetic properties, theresults suggest that Compounds 30.1 and 31.5 are likely to reduce viraltiters in human patients.

Example H Liver NTP Generation Following Oral Administration ofNucleoside Analogues and their Prodrugs

The potential for oral efficacy of various nucleosides and prodrugs wasassessed in the rat by evaluating liver NTP generation following oraladministration.

Methods:

Nucleoside analogues and their prodrugs were administered at 30 mg/kg(in terms of nucleoside equivalents) to Sprague-Dawley rats by oralgavage. At 2 or 3 hours following drug administration, liver samples (˜1g) were collected, snap-frozen, and homogenized in 3 volumes of ice-cold60% acetonitrile. Following centrifugation to clarify the homogenate,aliquots of the supernatants (100 μL) were evaporated to dryness on aSavant Speed-Vac Plus (1 hr, room temperature). The resulting driedresidue was reconstituted with 100 μL of mobile phase and then analyzedfor nucleotides by an LC-MS/MS method as described below.

The reconstituted extracts in mobile phase A (20 mMN,N-dimethylhexylamine and 10 mM propionic acid in 20% methanol) wereanalyzed by LC-MS/MS (Applied Biosystems, API 4000) equipped with anAgilent 1100 binary pump and a LEAP injector. Ten μL of sample wasinjected onto an Xterra MS C18 column (3.5 um, 2.1×50 mm, Waters Corp.)with a SecurityGuard C18 guard column (5 um, 4.0×3.0 mm, Phenomenex) andeluted with a gradient mobile phase A and B (20 mMN—N-dimethylhexylamine and 10 mM propionic acid in 80% methanol) at aflow rate of 0.3 mL/min (0 min, 0% B, 0-1 min, 0-50% B; 1-3 min, 50-100%B, 3-6 min, 100% B; 6-6.1 min, 100-0% B; 6.1-9 min, 0% B). NTP wasdetected by using MS/MS mode (M⁻/78.8). If nucleotide standards wereavailable, then the quantitative analysis of liver NTP was calculatedbased on a calibration curve generated from their respective standards(0.01, 0.03, 0.1, 0.3, 1.0, 3, 10, and 30 μM).

Results:

Liver NTP Dose, at 2 or 3 h, No. mg/kg nmoles/g 23.1 50 8.19 23.2 506.83 30.1 50 28.28 30.2 50 29.17 30.3 50 22.87 30.4 50 14.65 30.5 506.13 30.6 50 7.17 30.7 50 11.84 31.5 50 460 30.8 50 11.21 30.9 50 6.8730.1 50 20.27 30.11 50 2.68 30.12 50 6.8 30.13 50 2.17 30.14 50 5.0430.15 50 4.75 34.2 50 <0.4 31.3 8 0.02 36.1 3.5 28.89 34.3 50 <0.4 36.210 3.66

Conclusions:

The majority of the compounds of the invention evaluated generated thecorresponding NTP in liver following oral administration to rats. Thisindicates that the compounds have good oral bioavailability and, as HCVis a hepatotrophic virus, have the potential of demonstrating antiviralactivity in vivo.

Example I Assessment of Oral Bioavailability of 2′,3′-Carbonate Prodrugsof 2′-B-C-methyladenosine in the Rat Based on Liver NucleotideConcentrations

The oral bioavailability (OBAV) of 2′-β-C-methyladenosine and itscarbonate prodrugs was evaluated in the rat based on the liverconcentration-time profile of the generated nucleotides, including the5′-triphosphate (NTP), the form of the nucleoside that is generallyconsidered the active antiviral agent.

Methods:

2′-β-C-Methyladenosine and its carbonate prodrugs, Compound 31.5,Compound 30.1, and Compound 30.10, were administered by IV bolus andoral gavage to separate sets of male Sprague-Dawley rats. Atpre-specified times of 20 min, 1, 3, 5, 8, 16, and 24 hrs post dose,liver samples (˜1 g) were collected by freeze-clamping and homogenizedin 3 volumes of ice-cold 60% acetonitrile. Following centrifugation toclarify the homogenate, aliquots of the supernatants (100 μL) wereevaporated to dryness on a Savant Speed-Vac Plus (1 hr, roomtemperature). The resulting dried residue was reconstituted with 100 μLof mobile phase and then analyzed for nucleotides by an LC-MS/MS methodas described below.

The reconstituted extracts in mobile phase A (20 mMN,N-dimethylhexylamine and 10 mM propionic acid in 20% methanol) wereanalyzed by LC-MS/MS (Applied Biosystems, API 4000) equipped with anAgilent 1100 binary pump and a LEAP injector. Ten μL of sample wasinjected onto an Xterra MS C18 column (3.5 um, 2.1×50 mm, Waters Corp.)with a SecurityGuard C18 guard column (5 um, 4.0×3.0 mm, Phenomenex) andeluted with a gradient mobile phase A and B (20 mMN,N-dimethylhexylamine and 10 mM propionic acid in 80% methanol) at aflow rate of 0.3 mL/min (0 min, 0% B, 0-1 min, 0-50% B; 1-3 min, 50-100%B, 3-6 min, 100% B; 6-6.1 min, 100-0% B; 6.1-9 min, 0% B). NMP, NDP, andNTP were detected by using MS/MS mode (M/78.8). Where possible, thequantitative analysis of liver NMP, NDP, and NTP were calculated basedon a calibration curve generated from their respective standards(0.01-30 μM).

The area under the curve (AUC) to the last measurable time point werecalculated by trapezoidal summation of the liver concentration-timeprofile of total nucleotides and NTP. The OBAV was determined bydividing the dose-normalized AUC following oral administration by theAUC following IV dosing.

Results:

The OBAV values of 2′-β-C-methyladenosine and its carbonate prodrugs,Compounds 31.5, 30.1, and 30.10 in the rat are summarized in the tablebelow. The OBAV of the free nucleoside is very low (<5%) whereas theOBAV of its carbonate prodrugs are >20%.

ORAL BIOAVAILABILITY COMPOUND (%) 2′-β-C-Methyladenosine 2 31.5 95 30.139  30.10 21

Conclusions:

The OBAV of 2′-β-C-methyladenosine, was significantly enhanced as a2′,3′-carbonate prodrug. The presence of the carbonate moiety improvesthe OBAV potentially by increasing permeability of the nucleoside in thegut and/or by protecting the nucleoside from non-productive metabolism(see Example E).

Example J Evaluation of Liver Selectivity of an Antiviral Nucleoside

Analog and Its 2′,3′-Carbonate Prodrugs The liver selectivity of anantiviral nucleoside, 2′-B-C-methyladenosine, and its 2′,3′-carbonateprodrugs was evaluated in the rat by comparing liver nucleotideconcentrations to levels of a major metabolite in the plasma or toconcentrations of extrahepatic nucleotides. An elevated liverselectivity index is an indication of improved liver targeting of theantiviral agent and reduced exposure to other organs that may bepotential targets of toxicity.

Methods:

2′-β-C-Methyladenosine and its carbonate prodrugs, Compound 31.5,Compound 30.1, and Compound 30.10, were administered by IV bolus andoral gavage to separate sets of male Sprague-Dawley rats. Atpre-specified times of 20 min, 1, 3, 5, 8, 16, and 24 hrs post dose,liver, heart, and thigh muscle samples (˜1 g) were harvested byfreeze-clamping and homogenized in 3 volumes of ice-cold acetonitrile.Following centrifugation to clarify the homogenate, aliquots of thetissue extracts were evaporated to dryness on a Savant Speed-Vac Plus (1hr, room temperature). The resulting dried residue was reconstitutedwith 100 μL of mobile phase and then analyzed for nucleotides by anLC-MS/MS method as described in Example I. In addition to solid tissue,blood was collected in heparin tubes at the pre-specified times andcentrifuged to obtain plasma. The plasma samples (100 μL) were extractedwith 1.5 volumes of methanol and centrifuged for 20 min at top speed ona microfuge. Plasma concentrations of a major metabolite of2′-B-C-methyladenosine, i.e., 2′-B-C-methylinosine, were determined byLC-UV as described in Example E.

The area under the curve (AUC) to the last measurable time point wascalculated by trapezoidal summation of the liver, heart, and muscleconcentration-time profile of total nucleotides and NTP and of theplasma concentration-time profile of the major metabolite2′-B-C-methylinosine. The liver selectivity index was determined by twomethods: (1) dividing liver NTP AUC by heart nucleotide AUC and (2)dividing liver NTP AUC by plasma 2′-3-C-methylinosine AUC.

Results:

Whereas concentrations of nucleotides of 2′-methyladenosine weremeasurable in heart and muscle tissue following administration of thefree nucleoside, the nucleotides were barely detectable in these tissuesfollowing dosing of the 2′,3′-carbonate prodrugs. Plasma concentrationsof the main metabolite, 2′-B-C-methylinosine, were detectable afteradministration of the free nucleoside but not after dosing of theprodrug. A measure of liver targeting, the liver selectivity indices aretherefore significantly higher for the 2′,3′-carbonate prodrugs assummarized in the table below.

SELECTIVITY SELECTIVITY COMPOUND INDEX* INDEX** 2′-β-C- —  15/11 = 1.4Methyladenosine 31.5 2522/23 = 110 2522/25 = 101 30.1  841/2.6 = 323 841/<7.1 = >118  30.10  221/<2.8 = >75  211/<7.1 = >30 *Liver NTPAUC/Heart Nucleotides AUC **Liver NTP AUC/Plasma 2′-β-C-MethylinosineAUC

CONCLUSIONS

The delivery of the antiviral nucleoside analog, 2′-B-C-methyladenosine,to the liver by its carbonate prodrug not only led to an increase of thetherapeutic agent in the liver but also reduced peripheral drug exposureand any consequential toxicities. By diminishing the release ofpotential toxic elements to the systemic circulation, hepatic extractionand/or metabolism improved the liver targeting index of the nucleosideanalog.

1-29. (canceled)
 30. A compound of Formula I:

or an isomer, solvate, hydrate, prodrug, or pharmaceutically acceptablesalt thereof, wherein: X′ is O, S, S—O, or NR²⁰, wherein R²⁰ is H oroptionally substituted alkyl, aryl, arylalkyl, C₃₋₆ cycloalkyl, OH,OR^(20′), or O(C═O)R^(20′), wherein R^(20′) is H, lower alkyl or C₃₋₆cycloalkyl; Y is —O—, —S—, —N—, —C(R^(20′))—, or —CH₂—; R¹⁹ is H oroptionally substituted C₁₋₄ alkyl, C₂₋₄ alkenyl, or C₂₋₄ alkynyl, —OH,—O-lower alkyl, halogen, CN, or —C═CR²¹R²², wherein R²¹ and R²² areindependently H or lower alkyl; or R¹⁹ is absent; or R¹⁹ is joinedtogether with R¹⁷ to form —(CH₂)_(p)—, —O—(CH₂)_(p)—, wherein p is 0 to4; R¹⁸ is independently H, C₁₋₄ alkyl, C₂₋₄ alkenyl, or C₂₋₄ alkynyl;wherein said C₁₄ alkyl is optionally substituted with amino, hydroxy, or1 to 3 fluorine atoms, C₁₋₄ alkylamino, dialkylamino, C₃₋₆cycloalkylamino, halogen, or alkoxy; R¹⁷ is H, halogen, alkyl optionallysubstituted with 1 to 3 fluorine atoms, C₁₋₁₀ alkoxy optionallysubstituted with C₁₋₃ alkoxy or 1 to 3 fluorine atoms, C₂₋₆ alkenyloxy,C₁₋₄ alkylthio, C₁₋₈ alkylcarbonyloxy, aryloxycarbonyl, azido, amino,alkylamino, or dialkyl amino; R¹⁶ and R¹⁵ are independently H, C₁₋₄alkyl, C₂₋₄ alkenyl, or C₂₋₄ alkynyl; wherein said C₁₋₄ alkyl isoptionally substituted with amino, hydroxy, or 1 to 3 fluorine atoms,and said C₂₋₄ alkenyl and C₂₋₄ alkynyl are each optionally substitutedwith one or more of C₁₋₃ alkoxy, carboxy, C₂₋₆ alkenyloxy, C₁₋₄alkylthio, C₁₋₈ alkylcarbonyloxy, aryloxycarbonyl, azido, amino,alkylamino, or dialkylamino; B is selected from

wherein: A, D, E, J, and G are each independently selected from thegroup consisting of C and N; L is selected from O or S; M is selectedfrom the group consisting of O, S, and Se; X₁ is absent, or X₁ isselected from the group consisting of H, —OH, —SH, —NH₂, —COOR¹¹,—CONH₂, —CSNH₂, alkylamino, dialkylamino, cycloalkylamino, halogen,alkyl, alkenyl, alkynyl, aryl, alkaryl, cycloalkyl, acyl, alkoxy, CF₃,and —NHCOR_(X1), wherein R_(X1) is H, lower alkyl, or lower alkoxy, andwherein R¹¹ is H or C₁₋₄ alkyl; X₂ is absent, or X₂ is independentlyselected from the group consisting of H, alkenyl, alkynyl, aryl,alkaryl, cycloalkyl, acyl, and C₁-C₆ alkyl; X₃, X₄ and X₆ are eachindependently absent, or X₃, X₄ and X₆ are each independently selectedfrom the group consisting of H, alkenyl, alkynyl, aryl, alkaryl,cycloalkyl, acyl, OH, SH, NH₂, CF₃, alkyl, amino, halogen, alkylamino,cycloalkylamino, and dialkylamino; wherein when B contains J and G, thenX₁ and X₃ cannot both be (a) X₁═NH₂, alkylamino, dialkylamino,cycloalkylamino or —HCOR_(X1); and (b) X₃═NH₂, amino, alkylamino,cycloalkylamino or dialkylamino; X₅ is absent, or X₅ is selected fromthe group consisting of H, —CN, —NO₂, -alkyl, alkenyl, alkynyl, aryl,alkaryl, cycloalkyl, acyl, —NHCONH₂, —CONR¹¹R^(11′), —CSNR¹¹R^(11′),—COOR¹¹, —C(═NH)NH₂, -hydroxy, —C₁₋₃alkoxy, -amino, -alkylamino,-dialkylamino, halogen, -(1,3-oxazol-2-yl), -(1,3-thiazol-2-yl), and-(imidazol-2-yl); wherein alkyl is unsubstituted or substituted with oneto three groups independently selected from halogen, amino, hydroxy,carboxy, and C₁₋₃ alkoxy; and wherein R¹¹ and R^(11′) are independentlyH or C₁₋₄ alkyl; Z′ is —CH(R²³)OH, —O—, —CH(R²³)—, C₁₋₄cycloalkyl,—OC(R²³)₂PO₃H₂, —CH₂C(R²³)₂PO₃H₂, C₂₋₄ alkenyl, or C₂₋₄ alkynyl; whereinR²³ is H, F, methyl, ethyl, hydroxymethyl, or fluoromethyl or —CH₂N₃,—CH₂—NR²¹R²², and R²¹ and R²² are as defined above; and Z″ is absent, orZ″ is R²⁴(C═O)—, R²⁴—O—(C═O)—, or R²⁴CH(NH₂)(C═O)—, wherein R²⁴ isoptionally substituted C₁₋₆ alkyl, cycloalkyl, aryl, or aralkyl; or Z″is

wherein: V, W, and W′ are independently H, optionally substituted alkyl,optionally substituted aralkyl, cycloalkyl, heterocycloalkyl, aryl,substituted aryl, heteroaryl, substituted heteroaryl, optionallysubstituted 1-alkenyl, or optionally substituted 1-alkynyl; and Z is—CHR^(z)OH, —CHR^(z)OC(O)R^(y), —CHR^(z)OC(S)R^(y), —CHR^(z)OC(S)OR^(y),—CHR^(z)OC(O)SR^(y), —CHR^(z)OCO₂R^(y), —OR^(z), —SR^(z), —CHR^(z)N₃,—CH₂aryl, —CH(aryl)OH, —CH(CH═CR^(z) ₂)OH, —CH(C≡CR^(z))OH, —R^(z),—NR^(z) ₂, —OCOR^(y), —OCO₂R^(y), —SCOR^(y), —SCO₂R^(y), —NHCOR^(z),—NHCO₂R^(y), —CH₂NHaryl, —(CH₂)_(q)—OR^(z), and —(CH₂)_(q)—SR^(z),halogen, —CN, —COR^(y), —CONR^(z) ₂, —CO₂R^(y), —SO₂R^(y), or —SO₂NR^(z)₂, wherein q is 2 or 3, R^(z) is R^(y) or —H, and R^(y) is alkyl, aryl,cycloalkyl, heterocycloalkyl, or aralkyl; or Z″ is P(O)Y′R¹¹Y″R¹¹;wherein each R¹¹ is independently H; Y′ and Y″ are each independentlyselected from the group consisting of —O—, and —NR^(v)—; and when Y′ andY″ are both —O—, R¹¹ attached to —O— is independently selected from thegroup consisting of optionally substituted aryl, optionally substitutedCH₂-heterocycloakyl wherein the cyclic moiety contains a carbonate orthiocarbonate, optionally substituted -alkylaryl, —C(R^(z))₂OC(O)NR^(z)₂, —NR^(z)—C(O)—R^(y), —C(R^(z))₂—OC(O)R^(y), —C(R^(z))₂—O—C(O)OR^(y),—C(R^(z))₂OC(O)SR^(y), -alkyl-S—C(O)R^(y), -alkyl-S—S-alkylhydroxy, and-alkyl-S—S—S-alkylhydroxy; or when Y′ and Y″ are both —NR^(v)—, then R¹¹attached to —NR^(v)— is independently selected from the group consistingof —H, —[C(R^(z))₂]_(q)—COOR^(y), —C(R^(x))₂COOR^(y),—[C(R^(z))₂]_(q)—C(O)SR^(y), and -cycloalkylene-COOR^(y); or when Y′ is—O— and Y″ is NR^(v), then R¹¹ attached to —O— is independently selectedfrom the group consisting of optionally substituted aryl, optionallysubstituted CH₂-heterocycloakyl wherein the cyclic moiety contains acarbonate or thiocarbonate, optionally substituted -alkylaryl,—C(R^(z))₂OC(O)NR^(z) ₂, —NR^(z)—C(O)—R^(y), —C(R^(z))₂—OC(O)R^(y)—,—C(R^(z))₂—O—C(O)OR^(y), —C(R^(z))₂OC(O)SR^(y), -alkyl-S—C(O)R^(y),-alkyl-S—S-alkylhydroxy, and -alkyl-S—S—S-alkylhydroxy; and R¹¹ attachedto —NR^(v)— is independently selected from the group consisting of —H,—[C(R^(z))₂]_(q)—COOR^(y), —C(R^(x))₂COOR^(y),—[C(R^(z))₂]_(q)—C(O)SR^(y), and -cycloalkylene-COOR^(y); or when Y′ andY″ are independently selected from —O— and —NR^(v)—, then R¹¹ and R¹¹together form a cyclic group comprising -alkyl-S—S-alkyl-, wherein q isan integer 2 or 3; each R^(z) is selected from the group consisting ofR^(y) and —H; each R^(y) is selected from the group consisting of alkyl,aryl, heterocycloalkyl, and aralkyl; each R^(x) is independentlyselected from the group consisting of —H, and alkyl, or together R^(x)and R^(x) form a cycloalkyl group; and each R^(v) is selected from thegroup consisting of —H, lower alkyl, acyloxyalkyl,alkoxycarbonyloxyalkyl, and lower acyl; with the provisos that: a) V, Z,W, W′ are not all —H; b) when Z is —Rz, then at least one of V, W, andW′ is not —H, alkyl, aralkyl, cycloalkyl, or heterocycloalkyl; c) whenZ₁ is —CH₂OH and R⁷ is H, then one of R¹⁵, R¹⁶, R¹⁷ and R¹⁸ is otherthan H; and d) when Z′ is —CH₂O—, Z″ is —(C═O)R²⁴, and R¹⁷ is H, thenone of R¹⁵, R¹⁶, R¹⁷ and R¹⁸ is other than H.
 31. The compound of claim30, wherein Z″ is absent, or Z″ is R²⁴(C═O)—, R²⁴—O—(C═O)—, orR²⁴CH(NH₂)(C═O)—, wherein R²⁴ is optionally substituted C₁₋₆ alkyl,cycloalkyl, aryl, or aralkyl.
 32. The compound of claim 32, wherein X′is O, Y is O, and R¹⁹ is absent.
 33. The compound of claim 31, whereinX′ is S.
 34. The compound of claim 31, wherein R¹⁶ is —CH₃.
 35. Thecompound of claim 32, wherein Z″ is absent, Z′ is —CH(R²³)OH, R¹⁶ isC₁₋₄ alkyl, and R¹⁵ is H or C₁₋₄ alkyl.
 36. The compound of claim 35,wherein said compound is selected from the group consisting of:


37. The compound of claim 30, wherein: V and Z are connected togethervia an additional 3-5 atoms to form a cyclic group containing 5-7 atoms,wherein 0-1 atoms are heteroatoms and the remaining atoms are carbonsubstituted with hydroxy, acyloxy, alkylthiocarbonyloxy,alkoxycarbonyloxy, or aryloxycarbonyloxy attached to a carbon atom thatis three atoms from both 0 groups attached to the phosphorus; or V and Zare connected together via an additional 3-5 atoms to form a cyclicgroup, wherein 0-1 atoms are heteroatoms and the remaining atoms arecarbon, that is fused to an aryl group at the beta and gamma position tothe 0 attached to the phosphorus; or V and W are connected together viaan additional 3 carbon atoms to form an optionally substituted cyclicgroup containing 6 carbon atoms and substituted with one substituentselected from the group consisting of hydroxy, acyloxy,alkoxycarbonyloxy, alkylthiocarbonyloxy, and aryloxycarbonyloxy,attached to one of said carbon atoms that is three atoms from an Oattached to the phosphorus; or Z and W are connected together via anadditional 3-5 atoms to form a cyclic group, wherein 0-1 atoms areheteroatoms and the remaining atoms are carbon, and V must be aryl,substituted aryl, heteroaryl, or substituted heteroaryl; or W and W′ areconnected together via an additional 2-5 atoms to form a cyclic group,wherein 0-2 atoms are heteroatoms and the remaining atoms are carbon,and V must be aryl, substituted aryl, heteroaryl, or substitutedheteroaryl.
 38. A compound of Formula II′:

or an isomer, solvate, hydrate, prodrug, or pharmaceutically acceptablesalt thereof, wherein: X′ is O, S, S—O, or NR²⁰, wherein R²⁰ is H oroptionally substituted alkyl, aryl, arylalkyl, C₃₋₆ cycloalkyl, OH,OR^(20′), or O(C═O)R^(20′), wherein R^(20′) is H, lower alkyl or C₃₋₆cycloalkyl; Y is —O—, —S—, —N—, —C(R²⁰)—, or —CH₂—R¹⁹ is H or optionallysubstituted C₁₋₄ alkyl, C₂₋₄ alkenyl, or C₂₋₄ alkynyl, —OH, —O-loweralkyl, halogen, CN, or —C═CR²¹ R²², wherein R²¹ and R²² areindependently H or lower alkyl; or R¹⁹ is absent; or R¹⁹ is joinedtogether with R¹⁷ to form —(CH₂)_(p)—, —O—(CH₂)_(p)—, wherein p is 0 to4; R¹⁸ is independently H, C₁₋₄ alkyl, C₂₋₄ alkenyl, or C₂₋₄ alkynyl;wherein said C₁₋₄ alkyl is optionally substituted with amino, hydroxy,or 1 to 3 fluorine atoms, C₁₋₄ alkylamino, dialkylamino, C₃₋₆cycloalkylamino, halogen, or alkoxy; R¹⁷ is H, halogen, alkyl optionallysubstituted with 1 to 3 fluorine atoms, C₁₋₁₀ alkoxy optionallysubstituted with C₁₋₃ alkoxy or 1 to 3 fluorine atoms, C₂₋₆ alkenyloxy,C₁₋₄ alkylthio, C₁₋₈ alkylcarbonyloxy, aryloxycarbonyl, azido, amino,alkylamino, or dialkylamino; R¹⁶ and R¹⁵ are independently H, C₁₋₄alkyl, C₂₋₄ alkenyl, or C₂₋₄ alkynyl; wherein said C₁₋₄ alkyl isoptionally substituted with amino, hydroxy, or 1 to 3 fluorine atoms,and said C₂₋₄ alkenyl and C₂₋₄ alkynyl are each optionally substitutedwith one or more of C₁₋₃ alkoxy, carboxy, C₂₋₆ alkenyloxy, C₁₋₄alkylthio, C₁₋₈ alkylcarbonyloxy, aryloxycarbonyl, azido, amino,alkylamino, or dialkylamino; B is selected from the group consisting of:

wherein: A, D, E, J, and G are each independently selected from thegroup consisting of C and N; L is selected from O or S; M is selectedfrom the group consisting of O, S, and Se; X₁ is absent, or X₁ isselected from the group consisting of H, —OH, —SH, —NH₁₂, —COOR¹¹,—CONH₂, —CSNH₂, alkylamino, dialkylamino, cycloalkylamino, halogen,alkyl, alkoxy, CF₃, and —NHCOR_(X1), wherein R_(X1) is H, lower alkyl,or lower alkoxy, and wherein R¹¹ is H or C₁₋₄ alkyl; X₂ is absent, or X₂is independently selected from the group consisting of H and C₁₁C₆alkyl; X₃, X₄ and X₆ are each independently absent, or X₃, X₄ and X₆ areeach independently selected from the group consisting of H, OH, SH, NH₂,CF₃, alkyl, amino, halogen, alkylamino, cycloalkylamino, anddialkylamino; wherein when B contains J and G, then X₁ and X₃ cannotboth be (a) X₁═NH₂, alkylamino, dialkylamino, cycloalkylamino or—HCOR_(X1); and (b) X₃═NH₂, amino, alkylamino, cycloalkylamino ordialkylamino; X₅ is absent, or X₅ is selected from the group consistingof H, —CN, —NO₂, -alkyl, —NHCONH₂, —CONR¹¹R^(11′), —CSNR¹¹R^(11′),—COOR¹¹, —C(═NH)NH₂, -hydroxy, —C₁₋₃ alkoxy, -amino, -alkylamino,-dialkylamino, halogen, -(1,3-oxazol-2-yl), -(1,3-thiazol-2-yl), and-(imidazol-2-yl); wherein alkyl is unsubstituted or substituted with oneto three groups independently selected from the group consisting ofhalogen, amino, hydroxy, carboxy, and C₁₋₃ alkoxy; and wherein R¹¹ andR^(11′) are independently H or C₁₋₄ alkyl; Z′ is —O—, —CH(R²³)—O—, C₁₋₄cycloalkylene, C₂₋₄ alkenylene, or C₂₋₄ alkynylene; wherein R²³ ismethyl, ethyl, hydroxymethyl, fluoromethyl, —CH₂N₃, —CH₂—NR₂₁R₂₂; andR²¹ and R²² are as defined above; V, W, and W′ are independently H,optionally substituted alkyl, optionally substituted aralkyl,cycloalkyl, heterocycloalkyl, aryl, substituted aryl, heteroaryl,substituted heteroaryl, optionally substituted 1-alkenyl, or optionallysubstituted 1-alkynyl; and Z is —CHR^(z)OH, —CHR^(z)OC(O)R^(y),—CHR^(z)OC(S)R^(y), —CHR^(z)OC(S)OR^(y), —CHR^(z)OC(O)SR^(y),—CHR^(z)CO₂R^(Y), —OR^(z), —SR^(z), —CHR^(z)N₃, —CH₂aryl, —CH(aryl)OH,—CH(CH═CR^(z) ₂)OH, —CH(C≡CR^(z))OH, —R^(z), —NR^(z) ₂, —OCOR^(y),—OCO₂R^(y), —SCOR^(y), —SCO₂R^(y), —NHCOR^(z), —NHCO₂R^(y), —CH₂NHaryl,—(CH₂)_(q)—OR^(z), and —(CH₂)_(q)—SR¹, halogen, —CN, —COR^(y), —CONR^(z)₂, —CO₂R^(y), —SO₂R^(y), or —SO₂NR^(z) ₂, wherein q is 2 or 3, R^(z) isR^(y) or —H, and R^(y) is alkyl, aryl, cycloalkyl, heterocycloalkyl, oraralkyl; with the provisos that: a) V, Z, W, W′ are not all —H; and b)when Z is —Rz, then at least one of V, W, and W′ is not —H, alkyl,aralkyl, cycloalkyl, or heterocycloalkyl.
 39. The compound of claim 38,wherein X′ is O, Y is O, and R¹⁹ is absent.
 40. The compound of claim39, wherein R¹⁵, R¹⁶, R¹⁷, and R¹⁸ are independently H or C₁₋₄ alkyl.41. The compound of claim 38, wherein: V is selected from the groupconsisting of phenyl; substituted phenyl with 1-3 substituentsindependently selected from the group consisting of halogen, C₁₋₆ alkyl,—CF₃, —OR³, —OR¹², —COR³, —CO₂R³, —N(R³)₂, —N(R¹²)₂, —CO₂N(R²)₂, —SR³,—SO₂R³, —SO₂N(R²)₂ and —CN; monocyclic heteroaryl; and substitutedmonocyclic heteroaryl with 1-2 substituents independently selected fromthe group consisting of halogen, C₁₋₆ alkyl, —CF₃, —OR³, —OR¹², —COR³,—CO₂R³, —N(R³)₂, —N(R¹²)₂, —CO₂N(R²)₂, —SR³, —SO₂R³, —SO₂N(R²)₂ and —CN;wherein said monocyclic heteroaryl and substituted monocyclic heteroarylhas 1-2 heteroatoms that are independently selected from the groupconsisting of N, O, and S; wherein R² is H or R³, R³ is C₁₋₆ alkyl,aryl, heterocycloalkyl, or aralkyl, and R¹² is H or lower acyl; with theprovisos that a) when there are two heteroatoms and one is O, then theother can not be O or S, and b) when there are two heteroatoms and oneis S, then the other can not be O or S; or V and Z together areconnected via an additional 3-5 atoms to form a cyclic group, optionallycontaining I heteroatom, that is fused to an aryl group at the beta andgamma position to the O attached to the phosphorus.
 42. The compound ofclaim 41, wherein V is selected from the group consisting of phenyl;substituted phenyl with 1-2 substituents independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, and —CF₃; pyridyl;substituted pyridyl with 1 substituent independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, and —CF₃; furanyl;substituted furanyl with 1 substituent independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, and —CF₃; thienyl; andsubstituted thienyl with 1 substituent independently selected from thegroup consisting of —Cl, —Br, —F, C₁₋₃ alkyl, and —CF₃.
 43. The compoundof claim 38, wherein Z″ is:


44. The compound of claim 38, wherein Z″ is:


45. A pharmaceutical composition comprising a compound of claim 30, anda pharmaceutically acceptable excipient or carrier.
 46. A pharmaceuticalcomposition comprising a compound of claim 38, and a pharmaceuticallyacceptable excipient or carrier.
 47. A method of inhibiting viralreplication in a patient in need thereof, said method comprisingadministering to said patient a therapeutically effective amount of acompound of claim
 30. 48. The method of claim 47, wherein said viralreplication is RNA-dependent RNA viral replication.
 49. The method ofclaim 48, wherein said viral replication is HCV replication.
 50. Amethod of inhibiting viral replication in a patient in need thereof,said method comprising administering to said patient a therapeuticallyeffective amount of a compound of claim
 38. 51. The method of claim 50,wherein said viral replication is RNA-dependent RNA viral replication.52. The method of claim 51, wherein said viral replication is HCVreplication.
 53. A method of treating a viral infection in a patient inneed thereof, said method comprising administering to said patient atherapeutically effective amount of a compound of claim
 30. 54. Themethod of claim 53, wherein said viral infection is an RNA-dependent RNAviral infection.
 55. The method of claim 54, wherein said viralinfection is an HCV infection.
 56. The method of claim 55, wherein saidcompound is used in combination with a therapeutically effective amountof a second agent active against HCV.
 57. The method of claim 56,wherein said second agent active against HCV is ribavirin; levovirin;viramidine; thymosin alpha-1; interferon-β; an inhibitor of NS3 serineprotease; an inhibitor of inosine monophosphate dehydrogenase;interferon-α or pegylated interferon-α, alone or in combination withribavirin or levovirin.
 58. A method of treating a viral infection in apatient in need thereof, said method comprising administering to saidpatient a therapeutically effective amount of a compound of claim 38.59. The method of claim 58, wherein said viral infection is anRNA-dependent RNA viral infection.
 60. The method of claim 59, whereinsaid viral infection is an HCV infection.
 61. The method of claim 60,wherein said compound is used in combination with a therapeuticallyeffective amount of a second agent active against HCV.
 62. The method ofclaim 61, wherein said second agent active against HCV is ribavirin;levovirin; viramidine; thymosin alpha-1; interferon-β; an inhibitor ofNS3 serine protease; an inhibitor of inosine monophosphatedehydrogenase; interferon-α or pegylated interferon-α, alone or incombination with ribavirin or levovirin.
 63. A method of treatingcancer, liver fibrosis, diabetes, hyperlipidemia, obesity ornon-alcoholic steatohepatitis in a patient in need thereof, said methodcomprising administering to said patient a therapeutically effectiveamount of a compound of claim
 30. 64. A method of treating cancer, liverfibrosis, diabetes, hyperlipidemia, obesity or non-alcoholicsteatohepatitis in a patient in need thereof, said method comprisingadministering to said patient a therapeutically effective amount of acompound of claim
 38. 65. A method of treating a platelet disorder ordiabetes in a patient in need thereof, said method comprisingadministering to said patient a therapeutically effective amount of acompound of claim 30, wherein said compound is a P2 receptor antagonist.66. A method of treating a platelet disorder or diabetes in a patient inneed thereof, said method comprising administering to said patient atherapeutically effective amount of a compound of claim 38, wherein saidcompound is a P2 receptor antagonist.
 67. A method of treating diabetesin a patient in need thereof, said method comprising administering tosaid patient a therapeutically effective amount of a compound of claim30, wherein said compound is an AMPK activator.
 68. A method of treatingdiabetes in a patient in need thereof, said method comprisingadministering to said patient a therapeutically effective amount of acompound of claim 38, wherein said compound is an AMPK activator.
 69. Amethod of treating diabetes or cardiovascular disease in a patient inneed thereof, said method comprising administering to said patient atherapeutically effective amount of a compound of claim 30, wherein saidcompound binds an adenosine receptor.
 70. A method of treating diabetesor cardiovascular disease in a patient in need thereof, said methodcomprising administering to said patient a therapeutically effectiveamount of a compound of claim 38, wherein said compound binds anadenosine receptor.
 71. A method of treating inflammation or a CNSdisorder in a patient in need thereof, said method comprisingadministering to said patient a therapeutically effective amount of acompound of claim 30, wherein said compound acts as an adenosineanalogue.
 72. A method of treating inflammation or a CNS disorder in apatient in need thereof, said method comprising administering to saidpatient a therapeutically effective amount of a compound of claim 38,wherein said compound acts as an adenosine analogue.